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10.1101/gr.197152.115 http://scihub22266oqcxt.onion/10.1101/gr.197152.115 C4772021!4772021 !26813977     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=26813977 &cmd=llinks): Failed to open stream: HTTP request failed! 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A new class of temporarily phenotypic enhancers identified by CRISPR/Cas9-mediated genetic screening |pdf=|usr=}}{{26813977 }} gab.com Text A new class of temporarily phenotypic enhancers identified by CRISPR/Cas9-mediated genetic screening JO: Genome Res http://www.kidney.de/pget.php?&tw=1&pmid=26813977 #FOAvip With <2% of the human genome coding for proteins, a major challenge is to interpret the function of the noncoding DNA. Millions of regulatory sequences have been predicted in the human genome through analysis of DNA methylation, chromatin modification, hypersensitivity to nucleases, and transcription factor binding, but few have been shown to regulate transcription in their native contexts. We have developed a high-throughput CRISPR/Cas9-based genome-editing strategy and used it to interrogate 174 candidate regulatory sequences within the 1-Mbp POU5F1 locus in human embryonic stem cells (hESCs). We identified two classical regulatory elements, including a promoter and a proximal enhancer, that are essential for POU5F1 transcription in hESCs. Unexpectedly, we also discovered a new class of enhancers that contribute to POU5F1 transcription in an unusual way: Disruption of such sequences led to a temporary loss of POU5F1 transcription that is fully restored after a few rounds of cell division. These results demonstrate the utility of high-throughput screening for functional characterization of noncoding DNA and reveal a previously unrecognized layer of gene regulation in human cells. Twit Text FOAVip A new class of temporarily phenotypic enhancers identified by CRISPR/Cas9-mediated genetic screening ????Genome Res http://www.kidney.de/pget.php?&tw=1&pmid=26813977 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26813977 #FOAvip English Wikipedia <ref>{{Cite pmid|26813977 }}</ref> A new class of temporarily phenotypic enhancers identified by CRISPR/Cas9-mediated genetic screening #MMPMID26813977 Diao Y ; Li B ; Meng Z ; Jung I ; Lee AY ; Dixon J ; Maliskova L ; Guan KL ; Shen Y ; Ren B Genome Res 2016[Mar]; 26 (3 ): 397-405 PMID26813977 show ga With <2% of the human genome coding for proteins, a major challenge is to interpret the function of the noncoding DNA. Millions of regulatory sequences have been predicted in the human genome through analysis of DNA methylation, chromatin modification, hypersensitivity to nucleases, and transcription factor binding, but few have been shown to regulate transcription in their native contexts. We have developed a high-throughput CRISPR/Cas9-based genome-editing strategy and used it to interrogate 174 candidate regulatory sequences within the 1-Mbp POU5F1 locus in human embryonic stem cells (hESCs). We identified two classical regulatory elements, including a promoter and a proximal enhancer, that are essential for POU5F1 transcription in hESCs. Unexpectedly, we also discovered a new class of enhancers that contribute to POU5F1 transcription in an unusual way: Disruption of such sequences led to a temporary loss of POU5F1 transcription that is fully restored after a few rounds of cell division. These results demonstrate the utility of high-throughput screening for functional characterization of noncoding DNA and reveal a previously unrecognized layer of gene regulation in human cells. |*CRISPR-Cas Systems [MESH] |*Gene Targeting/methods [MESH] |*Genetic Testing/methods [MESH] |*Phenotype [MESH] |Cell Line [MESH] |Enhancer Elements, Genetic [MESH] |Gene Expression Regulation [MESH] |Genome, Human [MESH] |High-Throughput Screening Assays [MESH] |Humans [MESH] |Octamer Transcription Factor-3/genetics [MESH] |Regulatory Sequences, Nucleic Acid [MESH]A new class of temporarily phenotypic enhancers identified by CRISPR(epmc).pdf DeepDyve Pubget Overpricing