Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors
National Institute of Genetics · Research Organization of Information and Systems · +4 more institutions
Abstract
Studying the role of essential proteins is dependent upon a method for rapid inactivation, in order to study the immediate phenotypic consequences. Auxin-inducible degron (AID) technology allows rapid depletion of proteins in animal cells and fungi, but its application to human cells has been limited by the difficulties of tagging endogenous proteins. We have developed a simple and scalable CRISPR/Cas-based method to tag endogenous proteins in human HCT116 and mouse embryonic stem (ES) cells by using donor constructs that harbor synthetic short homology arms. Using a combination of AID tagging with CRISPR/Cas, we have generated conditional alleles of essential nuclear and cytoplasmic proteins in HCT116 cells,…
Citation impact
- FWCI
- 29.25
- Percentile
- 100%
- References
- 36
Authors
4- TNToyoaki Natsume
National Institute of Genetics, Research Organization of Information and Systems
- TKTomomi Kiyomitsu
Japan Science and Technology Agency, Nagoya University
- YSYumiko Saga
National Institute of Genetics, The Graduate University for Advanced Studies, SOKENDAI, The University of Tokyo
- MTMasato T. KanemakiCorresponding
National Institute of Genetics, The Graduate University for Advanced Studies, SOKENDAI, Japan Science and Technology Agency, Research Organization of Information and Systems
Topics & keywords
- Degron
- CRISPR
- Biology
- Cell biology
- Phenotype
- Endogeny
- Embryonic stem cell
- Cytoplasm