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The document discusses the identification of the hPRE-HoxA3 polycomb responsive element within the HoxA3 gene, which plays a critical role in regulating gene activity through long-range interactions with other genomic regions. The study utilizes various techniques, including qPCR, SiRNA construction, ChIP assays, and chromosome conformation capture, to analyze the regulatory functions of hPRE-HoxA3 in the context of Hox gene expression and embryonic development. Overall, the findings enhance the understanding of molecular mechanisms governing chromatin structure and gene regulation.

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6 views1 page

Summary Page.....

The document discusses the identification of the hPRE-HoxA3 polycomb responsive element within the HoxA3 gene, which plays a critical role in regulating gene activity through long-range interactions with other genomic regions. The study utilizes various techniques, including qPCR, SiRNA construction, ChIP assays, and chromosome conformation capture, to analyze the regulatory functions of hPRE-HoxA3 in the context of Hox gene expression and embryonic development. Overall, the findings enhance the understanding of molecular mechanisms governing chromatin structure and gene regulation.

Uploaded by

sonilalit999151
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Summary page

Identification of a polycomb responsive region in human Hox-A cluster


and its long-range interaction with poolycomb enriched genomic
region.

Polycomb and Trithorax group protein binds with specific DNA elements, polycomb response
element(PRE). PREs play a crucial role in regulating polycomb group protein to silence or repress gene
activity. These are well developed in Drosophila, while only a few PREs have been reported in human.
We have identified hPRE-HoxA3 in intron of HoxA3 gene. It is a part of Polycomb dependent gene;
regulatory network, it repress luciferase activity in a PCG-dependent manner and demonstrating
enrichment with repressive H3K27Me3 marks.
In this work we have analysed long-range interaction of hPRE-HoxA3 with other genomic loci to
understand its gene target.
First of all, we take cell media(HEK293T and Hela cells) and materials and then according to the paper
the author did this, we have done plasmid construction of gene of interest, and then we have done a
quantitative PCR (qPCR) to check expression of gene(seen). After, we have done SiRNA
construction(knockdown\knockout) for validation.
After SiRNA construction we have done CHIP(chromatin immunoprecipitation Assay) assay to check
which protein binding to our expressed gene ( to check protein-DNA\gene interaction).
We have done chromosome conformation capture assay(3C) to check chromatin-chromatin
interaction(to check binding region of DNA). For validation of 3C we have done PCR amplification to
identify genomic interaction.
PRE in human Hox cluster and it involved in long-range interactions with HoxA,B,C,D clusters. Hox-gene
provide development of body structure along head-tail axis during embryonic development. In this
paper,we have study role of hPRE-HoxA3 in 3D chromosomal Domain organisation, and its potential dual
function as a PRE and TRE. It also suggests us that hPRE-HoxA3 play a crucial role in regulation of Hox
genes and genetic interactions and it provide expression and impact on developmental processes.
In this paper, we find out regulatory role of hPRE-HoxA3; by discovery of a polycomb-responsive
element. hPRE-HoxA3 is situated within intron of HoxA3 gene. This element repress luciferase activity in
PCG-dependent manner. It also tells us that, involvement of hPRE_HoxA3 in 3D chromosome domain
organisation and its role in long range interaction, with the PCG enriched region of HoxA,B,C,D.
This paper contributes about understanding of molecular mechanism and chromatin structure and gene
regulation and long-range chromosomal interaction.

Thanks

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