期刊
CELL
卷 173, 期 3, 页码 665-+出版社
CELL PRESS
DOI: 10.1016/j.cell.2018.02.033
关键词
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资金
- Waitt Advanced Biophotonics Core Facility of the Salk Institute
- NIH-NCI CCSG [P30 014195]
- NINDS Neuroscience Core Grant [NS072031]
- Waitt Foundation
- Stem Cell
- NGS
- Razavi Newman Integrative Genomics and Bioinformatics
- Flow Cytometry Core Facilities of the Salk Institute
- Helmsley Trust
- NIH-NCI [CCSG P30 014195]
- Chapman Foundation
- Catharina Foundation Fellowship
- Howard Hughes Medical Institute Hanna H. Gray Fellows program
- Salk Women AMP
- Science Special Award
- NIH through the Office of the Director [5 DP5 OD021369-02]
- National Institutes on Aging [5 R21 AG056811-02]
- Helmsley Charitable Trust
- NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE [P30NS072031] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE ON AGING [R21AG056811] Funding Source: NIH RePORTER
- OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [DP5OD021369] Funding Source: NIH RePORTER
Class 2 CRISPR-Cas systems endow microbes with diverse mechanisms for adaptive immunity. Here, we analyzed prokaryotic genome and metagenome sequences to identify an uncharacterized family of RNA-guided, RNA-targeting CRISPR systems that we classify as type VI-D. Biochemical characterization and protein engineering of seven distinct orthologs generated a ribonuclease effector derived from Ruminococcus flavefaciens XPD3002 (CasRx) with robust activity in human cells. CasRx-mediated knockdown exhibits high efficiency and specificity relative to RNA interference across diverse endogenous transcripts. As one of the most compact single-effector Cas enzymes, CasRx can also be flexibly packaged into adeno-associated virus. We target virally encoded, catalytically inactive CasRx to cis elements of pre-mRNA to manipulate alternative splicing, alleviating dysregulated tau isoform ratios in a neuronal model of frontotemporal dementia. Our results present CasRx as a programmable RNA-binding module for efficient targeting of cellular RNA, enabling a general platform for transcriptome engineering and future therapeutic development.
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