Journal article

A saturating mutagenesis CRISPR-Cas9-mediated functional genomic screen identifies cis- and trans-regulatory elements of Oct4 in murine ESCs

Matthew C Canver, Pratibha Tripathi, Michael J Bullen, Moshe Olshansky, Yogesh Kumar, Lee H Wong, Stephen J Turner, Samuel Lessard, Luca Pinello, Stuart H Orkin, Partha Pratim Das

Journal of Biological Chemistry | AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC | Published : 2020

Abstract

Regulatory elements (REs) consist of enhancers and promoters that occupy a significant portion of the noncoding 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 ∼12-kb cis-region (cis-REs; CREs) of the Oct4 gene locu..

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Grants

Awarded by National Health and Medical Research Council (NHMRC) of Australia


Awarded by National Human Genome Research Institute (NHGRI)


Funding Acknowledgements

This work was supported by National Health and Medical Research Council (NHMRC) of Australia Grants APP1159461 and APP1182804 (to P. P. D.) and National Human Genome Research Institute (NHGRI) Career Development Award R00HG008399 and Genomic Innovator Award R35HG010717 (to L. P.). S. H. O. is an Investigator of the Howard Hughes Medical Institute (HHMI). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.