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• W2991321019 abstract "Abstract Regulatory elements (REs) consist of enhancers and promoters that occupy a significant portion of the non-coding genome and control gene expression programs either in –cis or in – trans . Putative REs have been identified largely based on their regulatory features (co-occupancy of ESC-specific transcription factors, enhancer histone marks and DNase hypersensitivity) in mouse embryonic stem cells (mESCs). However, less has been established regarding their regulatory functions in their native context. We deployed cis- and trans- regulatory elements scanning through saturating mutagenesis and sequencing (ctSCAN-SMS) to target elements within the ∼12kb cis -region ( Cis- REs; CREs) of the Oct4 gene locus, as well as genome-wide 2,613 high-confidence trans- REs (TREs), in mESCs. ctSCAN-SMS identified 10 CREs and 12 TREs, as novel candidate REs of the Oct4 gene in mESCs. Furthermore, deletions of these candidate REs confirmed that the majority of the REs are functionally active, and CREs are more active than TREs in controlling Oct4 gene expression. A subset of active CREs and TREs physically interact with the Oct4 promoter to varying degrees; specifically, a greater number of active CREs compared to active TREs, physically interact with the Oct4 promoter. Moreover, comparative genomics analysis reveals that more number of active CREs than active TREs are evolutionary conserved between mouse and primates, including human. Taken together, our study demonstrates the reliability and robustness of ctSCAN-SMS screening to identify critical REs, and investigate their roles in the regulation of transcriptional output of a target gene (in this case Oct4) in their native context." @default.
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• W2991321019 date "2019-11-22" @default.
• W2991321019 modified "2023-09-26" @default.
• W2991321019 title "A saturating mutagenesis CRISPR-Cas9 mediated functional genomic screen identifies<i>cis-</i>and<i>trans-</i>regulatory elements of<i>Oct4</i>in murine ESCs" @default.
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• W2991321019 doi "https://doi.org/10.1101/851683" @default.
• W2991321019 hasPublicationYear "2019" @default.
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