TY  - JOUR
A1  - Gama Brambila, Rodrigo
A1  - Chen, Jie
A1  - Dabiri, Yasamin
A1  - Tascher, Georg
A1  - Němec, Václav
A1  - Münch, Christian
A1  - Song, Guangqi
A1  - Knapp, Stefan
A1  - Cheng, Xinlai
T1  - A chemical toolbox for labeling and degrading engineered Cas proteins
T2  - JACS Au
N2  - The discovery of clustered regularly interspaced short palindromic repeats and their associated proteins (Cas) has revolutionized the field of genome and epigenome editing. A number of new methods have been developed to precisely control the function and activity of Cas proteins, including fusion proteins and small-molecule modulators. Proteolysis-targeting chimeras (PROTACs) represent a new concept using the ubiquitin-proteasome system to degrade a protein of interest, highlighting the significance of chemically induced protein-E3 ligase interaction in drug discovery. Here, we engineered Cas proteins (Cas9, dCas9, Cas12, and Cas13) by inserting a Phe-Cys-Pro-Phe (FCPF) amino acid sequence (known as the π-clamp system) and demonstrate that the modified CasFCPF proteins can be (1) labeled in live cells by perfluoroaromatics carrying the fluorescein or (2) degraded by a perfluoroaromatics-functionalized PROTAC (PROTAC-FCPF). A proteome-wide analysis of PROTAC-FCPF-mediated Cas9FCPF protein degradation revealed a high target specificity, suggesting a wide range of applications of perfluoroaromatics-induced proximity in the regulation of stability, activity, and functionality of any FCPF-tagging protein.
KW  - CRISPR/Cas9
KW  - genome editing
KW  - π-clamp
KW  - Cas9 inhibitor
KW  - PROTAC
KW  - chemical-induced proximity
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/62833
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-628338
SN  - 2691-3704
N1  - We thank DFG for the financial support (CH 1690/2-1 and 2-3). This work was supported in part by the LOEWE Center Frankfurt Cancer Institute funded by the Hessen State Ministry for Higher Education, Research and the Arts [III L 5-519/03/03.001-(0015)].
VL  - 1
IS  - 6
SP  - 777
EP  - 785
PB  - ACS Publications
CY  - Washington, DC
ER  -