| Title: |
HiHo-AID2: boosting homozygous knock-in efficiency enables robust generation of human auxin-inducible degron cells |
| Authors: |
Shiqian Li, Yafei Wang, Miesje van der Stoel, Xin Zhou, Shrinidhi Madhusudan, Kristiina Kanerva, Van Dien Nguyen, Nazli Eskici, Vesa M. Olkkonen, You Zhou, Taneli Raivio, Elina Ikonen |
| Source: |
Genome Biology, Vol 25, Iss 1, Pp 1-26 (2024) |
| Publisher Information: |
BMC, 2024. |
| Publication Year: |
2024 |
| Collection: |
LCC:Biology (General)
LCC:Genetics |
| Subject Terms: |
Auxin-inducible degron 2, Homology-directed repair enhancers, Homozygous knock-in, Functional inactivation, Chemogenetics, Human embryonic stem cell (hESC), Biology (General), QH301-705.5, Genetics, QH426-470 |
| More Details: |
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. |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
1474-760X |
| Relation: |
https://doaj.org/toc/1474-760X |
| DOI: |
10.1186/s13059-024-03187-w |
| Access URL: |
https://doaj.org/article/b386292dc07c45e88b124344bcaa5ff8 |
| Accession Number: |
edsdoj.b386292dc07c45e88b124344bcaa5ff8 |
| Database: |
Directory of Open Access Journals |
