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Abstract: cis-regulatory elements (CREs) orchestrate the spatiotemporal control of gene expression. The regulatory activity of CREs is typically assessed by reporter assays, in which CREs are studied outside their endogenous context. To circumvent this problem, we developed a CRISPR-Cas9 knockin approach to study CREs in a scar-free genomic context. Here, we describe the design, transfection, and screening protocol to insert CREs in mouse embryonic stem cells. Our strategy can provide important insights into the sufficiency of CREs for gene expression control.
Fuente: STAR Protocols, 2022, 3, 101492
Publisher: Elsevier
Year of publication: 2022
No. of pages: 18
Publication type: Article
DOI: 10.1016/j.xpro.2022.101492
ISSN: 2666-1667
Spanish project: MCIN/AEI /10.13039/501100011033
Publication Url: https://doi.org/10.1016/j.xpro.2022.101492
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TOMÁS PACHANO
ALVARO RADA IGLESIAS
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