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 wjpmr, 2022,8(4), 208-215 SJIF Impact Factor: 5.922
Review Article
Pankaj et al. WORLD JOURNAL OF PHARMACEUTICAL
World Journal of Pharmaceutical and Medical Research
AND MEDICAL RESEARCH ISSN 2455-3301
www.wjpmr.com Wjpmr

CRISPR CAS9: A NEW TECHNOLOGY TO MODIFY GENOME- A REVIEW

Pankaj Bhatt*1, Suruchi Singh2, Narjes Alfuraiji3, Ali E. Al-Snafi4

1
Department of Pharmaceutics, KIET Groups of Institutions (KIET School of Pharmacy), Muradnagar, Ghaziabad,
U.P, India.
2
Department of Pharmacology, Glocal University, Saharanpur, U.P, India.
3
Department of Pharmacology, College of Medicine, University of Karbala, Iraq.
4
Department of Pharmacology, College of Medicine, University of Thiqar- Iraq.

*Corresponding Author: Pankaj Bhatt

Department of Pharmaceutics, KIET Groups of Institutions (KIET School of Pharmacy), Muradnagar, Ghaziabad, U.P, India.

Article Received on 20/02/2022 Article Revised on 10/03/2022 Article Accepted on 30/03/2022

ABSTRACT
The simplest, cheapest and most valuable method of modifying the genome was CRISPR technology, which assists
the scientist in gathering the DNA chain and reforming its volume. CRISPR is a most significant prospective
implementation again genetic imperfection to develop and protect assortment from broadening of illness. Different
catalogue databases were previously published arrange, audit papers traverse, and systemic data completed in a
different CRISPR implementation. Look into various implements and effects of gene editing types of equipment
and other areas. Likewise, it helps to improve benefits and virtue of matter related to CRISPR Cas9 abbreviation
was CRISPR. The technology has been modifying the resistance mechanisms or unicellular organisms such as
bacteria. The organisms employ the CRISPR inferred RNA protein, which deducts the virus's capability to attack.
Di action can do to destroy DNA from invading by far apart. It permits the constituents to relocate monitor and the
changing of the genes. An extensive review of seventy-six papers involving research and review works was done to
let the work become quickly, and straightforward forward valuable and unique restriction Method of standard
modern and recently developed system to cure illness and scrutinise. Even though various current therapeutic
systems and different therapeutic system, were available, gene editing technology was available to cure cancer,
Ocular, AIDS, muscular dystrophy, tons illness, blood disorders, and others that reduce pain. Furthermore, they
intermingled with some particular side effects. To resolve the problem of drug delivery, and it shows efficacy by
overcoming the side effects; however, nevertheless, the famous and safety well-being have been the significant
hindrances again their Victorious location.

KEYWORDS: CRISPR, Cas9, Gene editing, Genetic defects, Genome engineering, Gene driving.

1. INTRODUCTION sequences discovered in bacteria and was discovered by

Archaein 1990 that contains several copies of virus
CRISPR is the type of technology for gene editing that
genes.[7-9] The various copy-edit methods were zinc
allows scientists to cut and paste and turn genes into
finger nuclease transcription activator-like effect or
DNA. It is concerned with the DNA destroying defence
nucleases and the CRISPR-related system Cas9 every
system that is found in some prokaryotic. CRISPR
time the DNA sequence more so. It comprises only one
dominates in gene editing. The first gene editing
guide RNA molecule whereby the earliest 20 of the
performed by CRISPR was calculated as 6.8. There are
nucleotide harmonises to the DNA target,[10,11] so Cas9
many exploded cases that are being done by CRISPR
every time produces DSBS at a specific position by the
since 2013. Short form for CRISPR is clustered regularly
time RNA existence in genomic DNA.[12]
interspaced short palindromic repeats. The nuclease
associated with CRISPRs is Cas9.[1-3] The 29-nucleotide
BSB originated from CAS9 and for the particular RNA,
repeat separated with a 32-nt spacer sequence was first
cellular DNA is found to mix with DSB for machine
shown in bacteria in 1987. Subsequently, 40% was found
repair.[13] If the error is found in the non-homologous end
in the sequence bacterial genome and 90% in archaea.[4]
joining DNA system used for this process, mistakes are
Many types of cash jeans are well deserved and are
also made and rectified and introduced, leading to
adjusted to repeated elements.[5] In 2005, the spacer
disruption of gene function.[14-18] The snip DNA
sequence was found in phage genomes, which was
fragment is stored between the palindromic CRISPR
considered the year of memorable CRISPR.[6] The full
sequence to preserve the available memory to prevent
form of CRISPR describes the group of nucleic acid

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 Pankaj et al. World Journal of Pharmaceutical and Medical Research

future infection from the previous viral strain.[18] In this, from attacking.[33] They replace LR after being re-infused
we will explain how CRISPR opens a new structure for in the patient with a higher ability to attack cancer
cancer research. cells.[34]

What the heck is CRISPR? CRISPR - A versatile tool for genome engineering
To understand CRISPR in a much better way, we need to CRISPR and CRISPR-associated proteins are grouped up
study about 1987, when Japanese scientists used to study to target the foreign viral DNA to the adaptive
E. coli bacteria, the first sequence found and repeated in immunity.[35] The two different Macanese RNAs -
the organism of DNA was discovered and repeated.[20,21] CRISPR targeting and trance activating RND - activate
The biological importance and role of the sequence are and help guide the CAS protein for binding viral DNA
not known.[22] As time passed, many more scientists have sequences claimed.[36-38] Likewise, proteins induce some
found about it in the same manner for other bacteria's single nucleotide base changes that Cas9 variants.[39]
DNA.[23] A proper name is given to this sequence that is
Clustered.[24] Regularly Interspaces Short Palindromic Design of SgRNAs and bio information resources
Repeats. However, the role of CRISPR is still not yet Like earlier techniques and technology, CRISPR is being
known till 2007.[25] By studying Streptococcus bacteria, it used as a genome editing tool spread throughout the
is used in yoghurt and plays a vital role.[26] Bacteria are scientific community.[40] It is being supported by
at constant hit from viruses and produce enzymes that Developing and designing through open bioinformatics
help fight it.[27] Different viruses arise when killing the resources and analysing that is done through CRISPR
virus; it scoops the leftover virus code and breaks it into associated experiments. Recently, there are different
tiny pieces. After which, it is store in the space of types of online tools available for RNAs related to the
CRISPR in the bacterium genome. CRISPR also works efficiency and quality being measured.[41] The system
like a devil for viruses and bacteria that use genetically that helps to find RNA focusing on its efficiency is
saved cells to defend themselves against the next calculated from extensive screening data and integration
attack.[28] A particular attack enzyme is produce called through nucleotide composition and its position within
CRISPR associated protein 9 (Cas9) at new viral the gene model.[42,43]
infection.[29]
Target discovery by CRISPR/CAS9 screens
How CRISPR is used An essential tool used for cancer therapy treatment
CRISPR-based technology has been used for tasks in discovers that a novel target is CRISPR.[44] For CRISPR
recent years, whether related to the killing drug-resistant to be generated, a large variety of Geneknock outs is
superbugs or removing the responsibility of gene for the required. The third requires a variety of practical steps
disease or creating a molecular recording device. and bioinformatics. For every target gene to be started,
Concerns about risks and ethics and ethical issues related the primary step is to predict the efficiency of
to their application were raised. At last of the year 2018, sgRNAs.[45,46] Different CRISPR/Cas9 screens
Chinese scientist He Jiankui said that he has been using technologies are used to identify between two functional
CRISPR that is being carried in both and are still given entities.It was found that in the genome CRISPR
for investigation.[30] separated pancreatic cell lines in E3 Ubiquitin ligase ring
finger protein 43, especially endangered to eliminate the
CRISPR application Wnt ligand (a comprehensive family of release
 CRISPR in gene therapy and medicine glycoproteins) receptor frizzle-5(FZD5). The standard
 Diseases CRISPR technology can cure that is being merged has been forecast since, in the
algorithm, the small synthetic interlink of dissimilar
Scientists are dealing with eight diseases. With the help CRISPR separation was conducted.[47]
of CRISPR Cas9, it has become the primary condition
required to treat revolutionary technology. Interrogation of the non-coding genome of cancer
CRISPR has proved to be a remarkable tool that is used
Cancer to interrogate non-coding elements. Three cancer-
The common primary disease associated with increased associated genes, Cullin 3, Neurofibromatosis type 1,
mortality that is rising throughout the world is cancer. and Neurofibromatosis type 2, help identify the enhancer
CRISPR application is made first on cancer.[31] The first region.[48] It is being performed for the genomic region of
advanced CRISPR clinical trial is being conducted in about 700 KB that surrounds the candidates' complete
China that can test the ability of gene editing tools that reading frame. CRISPR requires one more piece of
are useful for treating patients with the esophagus.[32] information to identify target genes.[49] For this reason,
The testing is being done at Hangzhou Cancer Hospital, the genomic region, despite sgRNA, shows both TP53
beginning with the extraction being done with the binding property and enhancer Markers. It was hitting off
patient's immune T cells. Using CRISPR says it is a required element result for the resistance of cells
arranged to replace the gene encoded with the protein against HRASG12V induced senescence. It is connected
called PD-one few tumours can bring together the to TP53. Multiple enhancers in the presence of Cyclin-
protein on the surface of immune cells and stop them dependent kinase inhibitor 1A (CDKN1A), which are

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Pankaj et al. World Journal of Pharmaceutical and Medical Research

found on editing, have been identified to result in the AIDS

down-regulation of Cyclin-dependent kinase inhibitor 1A CRISPR technology plays an important role in fighting
(CDKN1A) and due to which it can easily escape from against AIDS. One way is to remove the DNA from the
oncogene-induced senescence. Non-coding RNAs are hiding place to the DNA in the immune cells. This
another class of poorly genetic elements.In general, three method is useful for attacking the virus, which did not
different CAS9 techniques were systematically used to place an inactive form, making it suitable for maximum
detect the role of long code RNA in cancer cells. Zhu et therapies to fight against viruses. Another way through
al. used pairs of sgRNAs to delete 651 long noncoding which the virus can be removed is for resistance to HIV
RNA genomic sequences. Subsequently,51 non- infection. Few people from birth have resistance of HIV
codinglong RNAs were changing the Proliferation to a mutation in a gene that is particularly called CCR5,
phenotype after elimination.[50] which ensures that HIV is used as the entry point for
infecting cells. Not allowing the virus to hold it together
Blood disorders mutation helps to change the complete structure of the
The primary CRISPR trial was started in Europe and the protein. Last year in China, this method is found in a
US to treat the patient in February for beta-thalassemia controversial case. For editing the embryos to make them
and sickle cell infection. The two blood problems resistant to HIV infection, CRISPR-Cas9 plays an
influence oxygen transport in the blood. It involves the important role. The researchers and experimenters
immature microorganisms of the bone marrow of the thought among the community that the baby with
patient and the CRISPR innovation created by CRISPR CRISPR has a greater risk that they may die earlier. The
Therapeutics and vertex drug. It generates a vital consensus seems that more and more information and
oxygen-carrying protein, which brings oxygen together study are being required before using it for humans.[57]
better than in the adult form. Another type of blood
disorder in which CRISPR technology can work is Cystic fibrosis
hemophilia. The technology works with casebia Cystic fibrosis is responsible for some respiratory
employing CRISPR in which the tool for gene editing is problems. The person with this disorder has an age
being conveyed directly to the liver. limitation of only 40 years after being treated. By editing
the mutation that causes cystic fibrosis, CRISPR
Eye-related genetic diseases technology plays a vital role in finding the necessary
Treating genetic blindness CRISPR is known as the best foundation in the CFTR gene. Scientists have shown that
technology. The hereditary form of blindness, which is CRISPR can be induced in humans, found in patients
due to a particular mutation, makes it easier for CRISPR- with cystic fibrosis that fixes the disease mutation. The
Cas9 to target and modify a single gene.[51] The eye is an next procedure is to test it in humans, both in medicine
immune-privileged part of the body, so it is minimal and in CRISPR therapeutics.[58] There are many different
here, which is an advantage of Insight that shows the ways of mutation In the CFTR Jean for cystic fibrosis.
possibility that CRISPR ensures the reaction against it This means that different areas of CRISPR therapy are
that can drive and produce side effects. The main reason used to develop different genetic defects.[58]
for childhood birth blindness is Lebercon genital
amaurosis. The medicine that is continuously progressing Muscular dystrophy
for CRISPR therapy is Editasmedicine. For this, there In general, the money is that Duchenne muscular
was no treatment.[52] The company's primary purpose is dystrophy is responsible for mutations in the DMD
to ensure the light-sensitive cell's functioning so that diastrophic gene responsible for the suitable protein for
children do not lose their vision. The most important muscle contraction. Mainly the patient who suffers from
therapeutic target used in the case of diseases is this disease, there is a lot of muscle collapse, and there is
VEGFR2.[53] no possible cure for this disease, yet research done in
mice proves that CRISPR technology is used to correct
Liver-related genetic diseases many genetic mutations found in the disease. Many
The treatment option used to cure liver-related diseases researchers in the USA have found the new idea instead
like hereditary tyrosinemia is the correction of genes for of the previous one used by CRISPR technology to
the CRISPR system.[54] System delivery ofCAS9 mRNA remove or reduce 12 strategic mutation hotspot that
dealing with the C 12–200 lipid in peace and the HDR covers a maximum of 3000 different mutations
template dealing with an AAV vector help correct responsible for the muscular disease. A company named
approximately 6% of hepatocytes found in the mouse exonic therapeutics is responsible for developing this
model of human hereditary tyrosinemia.[55,56] Groups are method.[59]
being developed to deliver CRISPR components to the
liver. BAMEA-016B And TT3 are the two MPs used for Huntington’s disease
systematic delivery of CAS9-mRNA, and gRNA Are the The neurodegenerative condition that has a substantial
mice liver that allows the PCSK9 gene to edit and for the and powerful genetic component is Huntington's disease.
regulation of PCSK9 protein level.[56] When there is an abnormal repetition of DNA sequences
found in the Huntington gene, this disease is caused.
Treatment of this disease is complicated as it can be

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 Pankaj et al. World Journal of Pharmaceutical and Medical Research

dangerous for the brain. Scientists are looking for CRISPR has shown an important role. It also has an
methods and techniques to edit the gene tool to make it impact on crop breeding. The editing of genomes helps
easier and safer.[60] get the modification in a session that saves time and is
simple for the conventional breeding scheme. CRISPR
CRISPR in other fields also proves the best approach for pyramid breeding.[74]
CRISPR – A new tool for gene driving technology
CRISPR clustered regularly into space-short Palindromic CRISPR in de-extinction
repeats, an editing tool that was being developed in 2011. Even though an idea like this was brought from creative
In this tool, Arena helps bring the Indo nucleus to a science writing, it was not scientist that was running on.
specific target with ease. The Endonuclease Cleaves that Since to get back animal which wire varnished the initial
DNA targeted sites are triggering the cell for repairing prospects of pigeon passenger the moment of notes
the double-strand breaks. The repair process makes American green wood resident employee. CRISPR
changes found in the target site. techniques investigators schedule to initiate gene through
the gene of pigeon passenger to recent present time
For repairs, the cell mainly uses variants present in the comparable the stripe back and of pigeon, the hybrids
chromosome in a process called HDR. After repairing, will have been bread for many creations till DNA
the drive allele is copied to the wild type of chromosome, progeny resembles that the varnished genus of the initial
removing wild type DNA sequences with the genome.[61] creation of resuscitative pigeon was anticipated to
generate in the year of 2020.
CRISPR has expanded transgenic animal research
The CRISPR-CAS9technology that is being developed in More nutritious fish
2012 has changed entirely in genetic engineering. As The genesis customises Salman fish which raise two fold
CRISPR enables the editing of targeted genomes in a rapidly were standard are yield in Canada. When the
simple, efficient, and economical manner, the process of CRISPR arrived, gene-editing scientist would have
creating transgenic animals became simpler in the span promoted an investigation group, presently CRISPR-
of just a few years. Moreover, the commercial CAS9 was employed to produce purified Salman, which
availability of synthetic single-guide RNA reduced the allowed him to increase in number better and become
experimental time and improved the editing efficiency mota less likely to suffer from illness. Furthermore, it
compared to in vitro transcribed guide RNA. For halts the Salman reproduce sexually it control flee the
instance, Synthego Synthetic RNA shows high editing Aquaculture provision the investigators currently
efficiencies and a high germline transmission rate, which research the characteristic that would strengthen CRISPR
are critical factors for successfully generating transgenic use. We were so excited to make the genesis more
animals. The development of CRISPR based on comforts for the fish as well as resistance to disease.
nonengineering technology edits the gene in such a more Anna Marsilius head of analysis group, spoke to me that
straightforward way that creates transgenic mice in the the technology could be used also to emphasise the
lab. They are now available to guide and instruct the omega-3 parliament of the fish in order to make well-
researchers and scientists.[62] being and even more famous in the upcoming days.

Application of CRISPR/CAS9 in plant genome Eradicating pests

editing We used to maintain the spread of infectious disease;
The application of CRISPR in the plants is developing. even an animal species figure capable of doing so,
In Arabidopsis, a new plant with various genes affecting the biological community of interacting
consisting of ATFLS2, ATSPL4, ATPDS3, etc., they are organisms and their physical environment by using
targeted with the different mutational efficiencies CRISPR. The gene-editing technology employs to
ranging from 1.1% to 84.8% in the first generation.[63-69] generate gene-drive which providing the genetic
These are heritably found in a variety of generations and alteration would take over by all progeny open out all
a percentage rate of 79.4.[69] For modifying the two over the habitat numbers throughout the many creations.
genes, a single CRISPR with two different expressional A year later, scientist forum UK Imperial College
cassettes is being developed. In tobacco, CRISPR is London confirm that the technology can also help the
being joined together with version VIGS technology, and mosquito genus, which is in charge of transmission of
the internet to it causes morphological changes found in paludism (malaria).Which initiates gene once both
the compound leaves.[70-72] CRISPR helps in editing, and parents of them take it the process of lay in the egg by
its use has been developed from time to time.[73] female anopheles mosquitoes would have been halted in
the experiment with the lock-up mosquito was cleaned
CRISPR applications in crop genetic improvement up once number create later on gene drive has been
The CRISPR system is straightforward, adequate, initiated.[75]
efficient, simple, and highly specific, helping to produce
target events. However, it is an essential tool that plays a Clinical applications of CRISPR
crucial role in the modification of the genome in plants. Long before developing a gene-editing technique, this
For basic and higher levels of plant biology research, has been talked about editing the cell culture model’s

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 Pankaj et al. World Journal of Pharmaceutical and Medical Research

genes, organism, and humans.[76] Traditionally in this ACKNOWLEDGEMENT

gene therapy, the new genetic material edited is inserted
All authors were responsible for the study's concept, data
into somatic cells, after which the edited gene is shown
collection, writing of the article, and reviewing.
to be effective in curing disease by displaying its
therapeutic effect.[77] Gene therapy trials have been in
REFERENCES
place since the 1980s, but most experiments have been
conducted over the years due to somatic cells not 1. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib
reacting appropriately with genes, the mutual immune N, et al. Multiplex genome engineering using
response to the host, and changing genetic information CRISPR/Cas systems. Science, 2013; 339(6121):
have failed. Clinical trials first published in 2014 819–823.
confirmed the possibility of gene editing technique in the 2. Mali P, Yang L, Esvelt KM, Aach J, Guell M,
right direction but did not use the zinc finger technique. DiCarlo JE, et al. RNA-guided human genome
In this test, the T cell present in mice was targeted by engineering via Cas9. Science, 2013; 339(6121):
ZFN using the HIV co-receptor. This concept, as 823–826.
confirmed by a study done by the chain et al. We have 3. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna
been handed over the general judgment from this aimless JA, Charpentier E. A programmable dual-RNA-
review article to target mutagenesis and numerous gene- guided DNA endonuclease in adaptive bacterial
editing CRISPR employ in different sectors. Editing role immunity. Science, 2012; 337(6096): 816–821.
or swap gene pronouncements The CRISPR-Cas9 4. Ishino Y, Shinagawa H, Makino K, Amemura M,
genome editing technique can be employed via the Nakata A. Nucleotide sequence of the IAP gene,
different method through mixing it with one more gene responsible for alkaline phosphatase isozyme
editing technique the CRISPR/Cas9 technique is conversion Escherichia coli, and identification of the
fundamental and translational Cancer research was gene product. J Bacteriol. 1987; 169(12): 5429–
valuable since which has extreme utility in the upcoming 5433.
time. In the coming years, pooled CRISPR screening 5. Horvath P, Barrangou R. CRISPR/Cas, the immune
technology will bring forth an additional extensive set of system of bacteria and archaea. Science, 2010;
the critical gene, which will play a vital function again 327(5962): 167–170.
cancer cells. The time of genetic engineer in demand was 6. Jansen R, van Embden JDA, Gaastra W, Schouls
increasing rapidly for prophylactic of disease, and LM. Identification of genes that are associated with
therefore we should keep an eye on the extensive DNA repeats in prokaryotes. Mol Microbiol, 2002;
influence for this field to remain in genetics as a starting 43(6): 1565–1575.
point of understanding the therapeutic area of the gene.- 7. Bolotin A, Quinquis B, Sorokin A, Ehrlich SD.
Clustered regularly interspaced short palindrome
CONCLUSION repeats (CRISPRs) have spacers of
extrachromosomal origin. Microbiology, 2005;
This review article presented the general evolution from
151(Pt 8): 2551–2561.
random to targeted mutagenesis and various genes
8. Mojica FJM, Díez-Villaseñor C, García-Martínez J,
editing (CRISPR) used in different fields. CRISPR-Cas9
Soria E. Intervening sequences of regularly spaced
is based on CRISPR-associated proteins and these can
prokaryotic repeats derive from foreign genetic
perform genome editing functions or change gene
elements. J Mol Evol, 2005; 60(2): 174–182.
expression. The CRISPR-Cas9 genome editing technique
9. Pourcel C, Salvignol G, Vergnaud G. CRISPR
can be used via a different method by combining it with
elements in Yersinia pestis acquire new repeats by
another gene editing technique. The CRISPR / Cas9
preferential uptake of bacteriophage DNA and
technique in fundamental and translational cancer
provide additional evolutionary studies tools.
research was valuable, which has extreme utility in the
Microbiology, 2005; 151(Pt 3): 653–663.
upcoming time. In the coming years, pooled CRISPR
10. Mahfouz MM, Li L, Shamimuzzaman M, Wibowo
screening technology would bring forth an additional
A, Fang X, Zhu J-K. De novo-engineered
extensive set of the vital gene, which will play a vital
transcription activator-like effector (TALE) hybrid
function again cancer cell lines. By the time of genetic
nuclease with novel DNA binding specificity creates
engineering, demand was increasing rapidly for
double-strand breaks. Proc Natl Acad Sci USA.
prophylactic of disease; by then, we should keep an eye
2011; 108(6): 2623–2628.
on extensive influences for this field to remain in genetic
11. Mishra R, Joshi RK, Zhao K. Genome editing in
as starting point of human disease and comprehend the
rice: Recent advances, challenges, and future
therapeutic area of gene.
implications. Front Plant Sci., 2018; 9: 1361.
12. Wu X, Kriz AJ, Sharp, PA. Target specificity of the
Funding
CRISPR-Cas9 system. Quant Biol., 2014; 2(2): 59–
Non.
70.
13. Liu L, Fan X-D. CRISPR-Cas system: a powerful
Conflict of interest
tool for genome engineering. Plant Mol, Biol., 2014;
The authors declare no conflict of interest, financial or
85(3): 209–218.
otherwise.

www.wjpmr.com │ Vol 8, Issue 4, 2022. │ ISO 9001:2015 Certified Journal │ 212

Pankaj et al. World Journal of Pharmaceutical and Medical Research

14. Ma X, Zhu Q, Chen Y, Liu Y-G. CRISPR/Cas9 29. Messing SAJ, Ton-Hoang B, Hickman AB,
platforms for genome editing in plants: McCubbin AJ, Peaslee GF, Ghirlando R, et al. The
Developments and applications. Mol Plant, 2016; processing of repetitive extragenic palindromes: the
9(7): 961–974. structure of a repetitive extragenic palindrome
15. Char SN, Neelakandan AK, Nahampun H, Frame B, bound to its associated nuclease. Nucleic Acids Res.,
Main M, Spalding MH, et al. An Agrobacterium- 2012; 40(19): 9964–9979.
delivered CRISPR/Cas9 system high-frequency 30. Gupta RM, Musunuru K. Expanding the genetic
targeted mutagenesis in maise. Plant Biotechnol J., editing tool kit: ZFNs, TALENs, and CRISPR-Cas9.
2017; 15(2): 257–268. J Clin Invest., 2014; 124(10): 4154–4161.
16. Xie K, Minkenberg B, Yang Y. Boosting 31. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-
CRISPR/Cas9 multiplex editing capability with the Tieulent J, Jemal A. Global cancer statistics, 2012:
endogenous tRNA-processing system. Proc Natl Global Cancer Statistics, 2012. CA Cancer J Clin,
Acad Sci USA., 2015; 112(11): 3570–3575. 2015; 65(2): 87–108.
17. Lowder LG, Zhang D, Baltes NJ, Paul JW 3rd, Tang 32. Shine J, Dalgarno L. The 3’-terminal sequence of
X, Zheng X, et al. A CRISPR/Cas9 toolbox for Escherichia coli 16S ribosomal RNA:
multiplexed plant genome editing and transcriptional complementarity to nonsense triplets and ribosome
regulation. Plant Physiol, 2015; 169(2): 971–985. binding sites. Proc Natl Acad Sci USA., 1974; 71(4):
18. Miao J, Guo D, Zhang J, Huang Q, Qin G, Zhang X, 1342–1346.
et al. Targeted mutagenesis in rice using a CRISPR- 33. Wood EJ. Molecular cloning. A laboratory manual.
Cas system. Cell Res., 2013; 23(10): 1233–1236. Biochem Educ, 1983; 11(2): 82.
19. Ormond KE, Mortlock DP, Scholes DT, Bombard 34. Hsu PD, Lander ES, Zhang F. Development and
Y, Brody LC, Faucett WA, et al. Human germline applications of CRISPR-Cas9 for genome
genome editing. Am J Hum Genet. 2017; 101(2): engineering. Cell., 2014; 157(6): 1262–1278.
167–176. 35. Deveau H, Barrangou R, Garneau JE, Labonté J,
20. Shinagawa H, Makino K, Nakata A, Brenner S. Fremaux C, Boyaval P, et al. Phage response to
Regulation of the Pho regulon in Escherichia coli K- CRISPR-encoded resistance in Streptococcus
12. J Mol Biol., 1983; 168(3): 477–488. thermophilus. J Bacteriol, 2008; 190(4): 1390–400.
21. Sancar A, Kacinski BM, Mott DL, Rupp WD. 36. Deltcheva E, Chylinski K, Sharma CM, Gonzales K,
Identification of the uvrC gene product. Proc Natl Chao Y, Pirzada ZA, et al. CRISPR RNA
Acad Sci USA., 1981; 78(9): 5450–5454. maturation by trans-encoded small RNA and host
22. Ishino Y, Krupovic M, Forterre P. History of factor RNase III. Nature, 2011; 471(7340): 602–
CRISPR-Cas from encounter with a mysterious 607.
repeated sequence to genome editing technology. J 37. Gasiunas G, Barrangou R, Horvath P, Siksnys V.
Bacteriol [Internet]. 2018 [cited 2020 Nov 2]; Cas9-crRNA ribonucleoprotein complex mediates
200(7). Available from: specific DNA cleavage for adaptive immunity in
https://jb.asm.org/content/200/7/e00580-17. bacteria. Proc Natl Acad Sci USA., 2012; 109(39):
23. Barrangou R, Fremaux C, Deveau H, Richards M, E2579-2586.
Boyaval P, Moineau S, et al. CRISPR provides 38. DOE/Lawrence Berkeley National Laboratory.
acquired resistance against viruses in prokaryotes. Programmable DNA scissors found for bacterial
Science, 2007; 315(5819): 1709–1712. immune system. Science Daily [Internet]. 2012 Jun
24. Cong L, Zhang F. Genome engineering using 28. [cited 2020 Nov 2]; Available from:
CRISPR-Cas9 system. Methods Mol Biol., 2015; https://www.sciencedaily.com/releases/2012/06/120
1239: 197–217. 628193020.htm.
25. Nitta M, Goto M, Shibuya N, Okawa Y. A novel 39. Komor AC, Kim YB, Packer MS, Zuris JA, Liu DR.
protein with alkaline phosphatase and protease Programmable editing of a target base in genomic
inhibitor activities in Streptomyces hiroshimensis. DNA without double-stranded DNA cleavage.
Biol Pharm Bull, 2002; 25(7): 833–836. Nature, 2016; 533(7603): 420–424.
26. Lazdunski A. Peptidases and proteases of 40. Miller JH, editor. Experiments in Molecular
Escherichia coli and Salmonella typhimurium. Genetics. New York, NY: Cold Spring Harbor
FEMS Microbiol Lett., 1989; 63(3): 265–276. Laboratory Press, 1972.
27. Dassa J, Fsihi H, Marck C, Dion M, Kieffer- 41. Makino K, Shinagawa H, Amemura M, Nakata A.
Bontemps M, Boquet PL. A new oxygen-regulated Nucleotide sequence of the phoB gene, the positive
operon in Escherichia coli comprises the genes for a regulatory gene for the phosphate regulon of
putative third cytochrome oxidase and for pH 2.5 Escherichia coli K-12. J Mol Biol., 1986; 190(1):
acid phosphatase (appA). Mol Gen Genet, 1991; 37–44.
229(3): 341–352. 42. Hart T, Tong AHY, Chan K, Van Leeuwen J,
28. Piggot PJ, Sklar MD, Gorini L. Ribosomal Seetharaman A, Aregger M, et al. Evaluation and
alterations controlling alkaline phosphatase design of genome-wide CRISPR/SpCas9 knockout
isozymes in Escherichia coli. J Bacteriol, 1972; screens. G3 (Bethesda), 2017; 7(8): 2719–2727.
110(1): 291–299.

www.wjpmr.com │ Vol 8, Issue 4, 2022. │ ISO 9001:2015 Certified Journal │ 213

Pankaj et al. World Journal of Pharmaceutical and Medical Research

43. Doench JG, Fusi N, Sullender M, Hegde M, 57. Lander ES. The heroes of CRISPR. Cell., 2016;
Vaimberg EW, Donovan KF, et al. Optimised 164(1–2): 18–28.
sgRNA design to maximise activity and minimise 58. Ferreira R, David F, Nielsen J. Advancing
off-target effects of CRISPR-Cas9. Nat Biotechnol, biotechnology with CRISPR/Cas9: recent
2016; 34(2): 184–191. applications and patent landscape. J Ind Microbiol
44. Bäumlein H, Pustell J, Wobus U, Case ST, Kafatos Biotechnol, 2018; 45(7): 467–480.
FC. The 3’ ends of two genes in the Balbiani ring c 59. Liu H, Wang L, Luo Y. Blossom of CRISPR
locus of Chironomusthummi. J Mol Evol., 1986; technologies and applications in disease treatment.
24(1–2): 72–82. Synth Syst Biotechnol, 2018; 3(4): 217–228.
45. Meier JA, Zhang F, Sanjana NE. GUIDES: sgRNA 60. BozorgQomi S, Asghari A, Mojarrad M. An
design for loss-of-function screens. Nat Methods. overview of the CRISPR-based genomic- and
2017; 14(9): 831–832. epigenome-editing system: Function, applications,
46. Heigwer F, Zhan T, Breinig M, Winter J, and challenges. Adv Biomed Res., 2019; 8(1): 49.
Brügemann D, Leible S, et al. CRISPR library 61. McCarty NS, Graham AE, Studená L, Ledesma-
designer (CLD): software for multispecies design of Amaro R. Multiplexed CRISPR technologies for
single guide RNA libraries. Genome Biol., 2016; gene editing and transcriptional regulation. Nat
17(1): 55. Commun, 2020; 11(1): 1281.
47. Tao Y, Mis M, Blazer L, Ustav M Jnr, Steinhart Z, 62. Ethics and research integrity [Internet]. Ethics-and-
Chidiac R, et al. Tailored tetravalent antibodies integrity.org. [cited Nov 3]. Available from:
potently and specifically activate Wnt/Frizzled http://ethics-and-
pathways in cells, organoids and mice. Elife integrity.org/ethics/ethicsGenomeEditing.html,
[Internet], 2019; 8. Available from: 2020.
http://dx.doi.org/10.7554/eLife.46134 63. Li J-F, Norville JE, Aach J, McCormack M, Zhang
48. Sanjana NE, Wright J, Zheng K, Shalem O, D, Bush J, et al. Multiplex and homologous
Fontanillas P, Joung J, et al. High-resolution recombination-mediated genome editing in
interrogation of functional elements in the non- Arabidopsis and Nicotianabenthamiana using guide
coding genome. Science, 2016; 353(6307): 1545– RNA and Cas9. Nat Biotechnol, 2013; 31(8): 688–
1549. 691.
49. Korkmaz G, Lopes R, Ugalde AP, Nevedomskaya 64. Jiang W, Zhou H, Bi H, Fromm M, Yang B, Weeks
E, Han R, Myacheva K, et al. Functional genetic DP. Demonstration of CRISPR/Cas9/sgRNA-
screens for enhancer elements in the human genome mediated targeted gene modification in Arabidopsis,
using CRISPR-Cas9. Nat Biotechnol, 2016; 34(2): tobacco, sorghum and rice. Nucleic Acids Res.,
192–198. 2013; 41(20): e188.
50. Europepmc.org. [cited Nov 2]. Available from: 65. Fauser F, Schiml S, Puchta H. Both CRISPR/Cas-
https://europepmc.org/article/med/28976505, 2020. based nucleases and nickases can be used efficiently
51. Ahn CH, Ramya M, An HR, Park PM, Kim Y-J, Lee for genome engineering in Arabidopsis thaliana.
SY, et al. Progress and challenges in the Plant J., 2014; 79(2): 348–359.
improvement of ornamental plants by genome 66. Song G, Jia M, Chen K, Kong X, Khattak B, Xie C,
editing. Plants, 2020; 9(6): 687. et al. CRISPR/Cas9: A powerful tool for crop
52. Komor AC, Badran AH, Liu DR. CRISPR-based genome editing. Crop J., 2016; 4(2): 75–82.
technologies for the manipulation of eukaryotic 67. Hyun Y, Kim J, Cho SW, Choi Y, Kim J-S,
genomes. Cell., 2017; 169(3): 559. Coupland G. Site-directed mutagenesis in
53. Shah F, Khan Z, Iqbal A, Turan M, Olgun M. Arabidopsis thaliana using dividing tissue-targeted
Recent advances in grain crops research. Shah F, RGEN of the CRISPR/Cas system to generate
Khan Z, Iqbal A, Turan M, Olgun M, editors. Intech heritable null alleles. Planta, 2015; 241(1): 271–
Open, 2020. 284.
54. Yin H, Song C-Q, Dorkin JR, Zhu LJ, Li Y, Wu Q, 68. Feng Z, Mao Y, Xu N, Zhang B, Wei P, Yang D-L,
et al. Therapeutic genome editing by combined viral et al. Multigeneration analysis reveals the
and non-viral delivery of CRISPR system inheritance, specificity, and patterns of
components in vivo. Nat Biotechnol, 2016; 34(3): CRISPR/Cas-induced gene modifications in
328–333. Arabidopsis. Proc Natl Acad Sci USA., 2014;
55. Li B, Luo X, Deng B, Wang J, McComb DW, Shi 111(12): 4632–4637.
Y, et al. An orthogonal array optimisation of lipid- 69. Mao Y, Zhang H, Xu N, Zhang B, Gou F, Zhu J-K.
like nanoparticles for mRNA delivery in vivo. Nano Application of the CRISPR-Cas system for efficient
Lett., 2015; 15(12): 8099–8107. genome engineering in plants. Mol Plant., 2013;
56. Liu J, Chang J, Jiang Y, Meng X, Sun T, Mao L, et 6(6): 2008–2011.
al. Fast and efficient CRISPR/Cas9 genome editing 70. Kumar R, Kaur A, Pandey A, Mamrutha HM, Singh
in vivo enabled by bioreducible lipid and messenger GP. CRISPR-based genome editing in wheat: a
RNA nanoparticles. Adv Mater, 2019; 31(33): comprehensive review and future prospects. Mol
e1902575. Biol Rep., 2019; 46(3): 3557–3569.

www.wjpmr.com │ Vol 8, Issue 4, 2022. │ ISO 9001:2015 Certified Journal │ 214

 Pankaj et al. World Journal of Pharmaceutical and Medical Research

71. Gao J, Wang G, Ma S, Xie X, Wu X, Zhang X, et al.

CRISPR/Cas9-mediated targeted mutagenesis in
Nicotianatabacum. Plant Mol Biol., 2015; 87(1–2):
99–110.
72. Sun X, Hu Z, Chen R, Jiang Q, Song G, Zhang H, et
al. Targeted mutagenesis in soybean using the
CRISPR-Cas9 system. Sci Rep., 2015; 5(1): 10342.
73. Brooks C, Nekrasov V, Lippman ZB, Van Eck J.
Efficient gene editing in tomato in the first
generation using the clustered regularly interspaced
short palindromic repeats/CRISPR-associated9
system. Plant Physiol, 2014; 166(3): 1292–1297.
74. Tabebordbar M, Zhu K, Cheng JKW, Chew WL,
Widrick JJ, Yan WX, et al. In vivo gene editing in
dystrophic mouse muscle and muscle stem cells.
Science, 2016; 351(6271): 407–411.
75. Pickar-Oliver A, Gersbach CA. The next generation
of CRISPR-Cas technologies and applications. Nat
Rev Mol Cell Biol., 2019; 20(8): 490–507.
76. Wirth T, Parker N, Ylä-Herttuala S. History of gene
therapy. Gene, 2013; 525(2): 162–169.
77. Yu L, Tian X, Gao C, Wu P, Wang L, Feng B, et al.
Genome editing for the treatment of tumorigenic
viral infections and virus-related carcinomas. Front
Med, 2018; 12(5): 497–508.
78. Cornu TI, Mussolino C, Cathomen T. Refining
strategies to translate genome editing to the clinic.
Nat Med, 2017; 23(4): 415–423.

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