{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["14(12)"],"submitter":["Liu J"],"pubmed_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."],"journal":["Protein & cell"],"pagination":["874-887"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC10691850"],"repository":["biostudies-literature"],"pubmed_title":["CRISPR-assisted transcription activation by phase-separation proteins."],"pmcid":["PMC10691850"],"pubmed_authors":["Liu J","Li Z","Yang Y","Songyang Z","Cui K","Nong B","Tan W","Chen Y","Luo X","Liang P","Zhang P","Ma W"],"additional_accession":[]},"is_claimable":false,"name":"CRISPR-assisted transcription activation by phase-separation proteins.","description":"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.","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Dec","modification":"2025-05-18T10:39:13.205Z","creation":"2025-05-18T10:39:13.205Z"},"accession":"S-EPMC10691850","cross_references":{"pubmed":["36905356"],"doi":["10.1093/procel/pwad013"]}}