CRISPR-assisted transcription activation by phase-separation proteins
Protein & Cell
; (12): 874-887, 2023.
Article
in En
| WPRIM
| ID: wpr-1010762
Responsible library:
WPRO
ABSTRACT
The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms. Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient transcriptional activation. Here, we fused different phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) and observed robust increases in transcriptional activation efficiency. Notably, human NUP98 (nucleoporin 98) and FUS (fused in sarcoma) IDR domains were best at enhancing dCas9-VPR activity, with dCas9-VPR-FUS IDR (VPRF) outperforming the other CRISPRa systems tested in this study in both activation efficiency and system simplicity. dCas9-VPRF overcomes the target strand bias and widens gRNA designing windows without affecting the off-target effect of dCas9-VPR. These findings demonstrate the feasibility of using phase-separation proteins to assist in the regulation of gene expression and support the broad appeal of the dCas9-VPRF system in basic and clinical applications.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Transcriptional Activation
/
Gene Expression Regulation
/
CRISPR-Cas Systems
/
RNA, Guide, CRISPR-Cas Systems
Limits:
Humans
Language:
En
Journal:
Protein & Cell
Year:
2023
Type:
Article