Universität zu Köln

Daliri, Karim ORCID: 0000-0003-1750-7046 and Clement, Kendell (2025). Protocol for permanent gene repression by CRISPR-adenine base editing of promoter CCAAT motifs. STAR Protocols, 6 (3). pp. 1-16. Elsevier. ISSN 2666-1667

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Abstract

Here, we present a protocol to achieve permanent downregulation of gene expression by editing the CCAAT box in promoter regions using CRISPR-adenine base editors (ABEs). We outline steps for guide RNA (gRNA) design, transfection, genomic DNA extraction, Sanger sequencing, and gene expression quantifica- tion. The protocol is optimized for mammalian cell lines (e.g., NIH3T3). It allows for precise disruption of transcription factor binding site without double-strand breaks and offers a novel alternative to RNAi or CRISPR interference (CRISPRi). For complete details on the use and execution of this protocol, please refer to Daliri et al.

Item Type:	Article
Creators:	Creators Email ORCID ORCID Put Code Daliri, Karim UNSPECIFIED https://orcid.org/0000-0003-1750-7046 UNSPECIFIED Clement, Kendell UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-809377
Identification Number:	10.1016/j.xpro.2025.104075
Journal or Publication Title:	STAR Protocols
Volume:	6
Number:	3
Page Range:	pp. 1-16
Number of Pages:	16
Date:	19 September 2025
Publisher:	Elsevier
ISSN:	2666-1667
Language:	English
Faculty:	Faculty of Medicine
Divisions:	Faculty of Medicine > Physiologie und Pathophysiologie > Institut für Neurophysiologie
Subjects:	Medical sciences Medicine
['eprint_fieldname_oa_funders' not defined]:	Publikationsfonds UzK
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/80937