Targeted regulation of transcription in primary cells using CRISPRa and CRISPRi.

ABSTRACT: Targeted transcriptional activation or interference can be induced with the CRISPR-Cas9 system (CRISPRa/CRISPRi) using nuclease-deactivated Cas9 fused to transcriptional effector molecules. These technologies have been used in cancer cell lines, particularly for genome-wide functional genetic screens using lentiviral vectors. However, CRISPRa and CRISPRi have not yet been widely applied to ex vivo cultured primary cells with therapeutic relevance owing to a lack of effective and nontoxic delivery modalities. Here we develop CRISPRa and CRISPRi platforms based on RNA or ribonucleoprotein (RNP) delivery by electroporation and show transient, programmable gene regulation in primary cells, including human CD34⁺ hematopoietic stem and progenitor cells (HSPCs) and human CD3⁺

SUBMITTER: Jensen TI 

PROVIDER: S-EPMC8559706 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature