HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells

Li, Shiqian, Wang, Yafei, van der Stoel, Miesje, Zhou, Xin, Madhusudan, Shrinidhi, Kanerva, Kristiina, Nguyen, Van Dien, Eskici, Nazli, Olkkonen, Vesa M., Zhou, You ORCID: https://orcid.org/0000-0002-1743-1291, Raivio, Taneli and Ikonen, Elina 2024. HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells. Genome Biology 25 , 58. 10.1186/s13059-024-03187-w

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Abstract

Recent developments in auxin-inducible degron (AID) technology have increased its popularity for chemogenetic control of proteolysis. However, generation of human AID cell lines is challenging, especially in human embryonic stem cells (hESCs). Here, we develop HiHo-AID2, a streamlined procedure for rapid, one-step generation of human cancer and hESC lines with high homozygous degron-tagging efficiency based on an optimized AID2 system and homology-directed repair enhancers. We demonstrate its application for rapid and inducible functional inactivation of twelve endogenous target proteins in five cell lines, including targets with diverse expression levels and functions in hESCs and cells differentiated from hESCs.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Advanced Research Computing @ Cardiff (ARCCA) Medicine Systems Immunity Research Institute (SIURI)
Publisher:	BioMed Central
ISSN:	1474-760X
Date of First Compliant Deposit:	12 March 2024
Date of Acceptance:	14 February 2024
Last Modified:	10 Jun 2024 09:30
URI:	https://orca.cardiff.ac.uk/id/eprint/166560

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