Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs.

Kundert, Kale; Lucas, James E; Watters, Kyle E; Fellmann, Christof; Ng, Andrew H; Heineike, Benjamin M; Fitzsimmons, Christina M; Oakes, Benjamin L; Qu, Jiuxin; Prasad, Neha; Rosenberg, Oren S; Savage, David F; El-Samad, Hana; Doudna, Jennifer A; Kortemme, Tanja

Kundert, Kale; Lucas, James E; Watters, Kyle E; Fellmann, Christof; Ng, Andrew H; Heineike, Benjamin M; Fitzsimmons, Christina M; Oakes, Benjamin L; Qu, Jiuxin; Prasad, Neha; Rosenberg, Oren S; Savage, David F; El-Samad, Hana; Doudna, Jennifer A; Kortemme, Tanja.

Afiliação

Kundert K; Graduate Group in Biophysics, University of California San Francisco, San Francisco, CA, 94158, USA. kale@thekunderts.net.
Lucas JE; UC Berkeley - UCSF Graduate Program in Bioengineering, University of California San Francisco, San Francisco, CA, 94158, USA.
Watters KE; Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94704, USA.
Fellmann C; Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94704, USA.
Ng AH; UC Berkeley - UCSF Graduate Program in Bioengineering, University of California San Francisco, San Francisco, CA, 94158, USA.
Heineike BM; Bioinformatics Graduate Program, University of California San Francisco, San Francisco, CA, 94158, USA.
Fitzsimmons CM; Chemistry and Chemical Biology Graduate Program, University of California San Francisco, San Francisco, CA, 94158, USA.
Oakes BL; Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
Qu J; Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94704, USA.
Prasad N; Department of Medicine, University of California San Francisco, San Francisco, CA, 94158, USA.
Rosenberg OS; Department of Medicine, University of California San Francisco, San Francisco, CA, 94158, USA.
Savage DF; Department of Medicine, University of California San Francisco, San Francisco, CA, 94158, USA.
El-Samad H; Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA.
Doudna JA; Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, 94704, USA.
Kortemme T; Chan Zuckerberg Biohub, San Francisco, CA, 94158, USA.

Nat Commun ; 10(1): 2127, 2019 05 09.

Article em En | MEDLINE | ID: mdl-31073154

RESUMO

The CRISPR-Cas9 system provides the ability to edit, repress, activate, or mark any gene (or DNA element) by pairing of a programmable single guide RNA (sgRNA) with a complementary sequence on the DNA target. Here we present a new method for small-molecule control of CRISPR-Cas9 function through insertion of RNA aptamers into the sgRNA. We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands. Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.

Assuntos

Aptâmeros de Nucleotídeos/genética; Proteína 9 Associada à CRISPR/genética; Sistemas CRISPR-Cas/genética; Edição de Genes/métodos; RNA Guia de Cinetoplastídeos/genética; DNA/genética; Ligantes

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Texto completo: 1 Base de dados: MEDLINE Assunto principal: RNA Guia de Cinetoplastídeos / Aptâmeros de Nucleotídeos / Sistemas CRISPR-Cas / Edição de Genes / Proteína 9 Associada à CRISPR Idioma: En Revista: Nat Commun Assunto da revista: BIOLOGIA / CIENCIA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Estados Unidos

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