TY - JOUR
T1 - HiHo-AID2
T2 - boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells
AU - Li, Shiqian
AU - Wang, Yafei
AU - van der Stoel, Miesje
AU - Zhou, Xin
AU - Madhusudan, Shrinidhi
AU - Kanerva, Kristiina
AU - Nguyen, Van Dien
AU - Eskici, Nazli
AU - Olkkonen, Vesa M.
AU - Zhou, You
AU - Raivio, Taneli
AU - Ikonen, Elina
N1 - Publisher Copyright:
© The Author(s) 2024.
PY - 2024
Y1 - 2024
N2 - 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.
AB - 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.
KW - Auxin-inducible degron 2
KW - Chemogenetics
KW - Functional inactivation
KW - hESC-derived neurons
KW - Homology-directed repair enhancers
KW - Homozygous knock-in
KW - Human embryonic stem cell (hESC)
KW - 318 Medical biotechnology
KW - 11832 Microbiology and virology
KW - 1184 Genetics, developmental biology, physiology
U2 - 10.1186/s13059-024-03187-w
DO - 10.1186/s13059-024-03187-w
M3 - Article
C2 - 38409044
AN - SCOPUS:85186397627
SN - 1474-7596
VL - 25
JO - Genome Biology
JF - Genome Biology
IS - 1
M1 - 58
ER -