Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells

Ayal Hendel; Rasmus O. Bak; Joseph T. Clark; Andrew B. Kennedy; Daniel E. Ryan; Subhadeep Roy; Israel Steinfeld; Benjamin D. Lunstad; Robert J. Kaiser; Alec B. Wilkens; Rosa Bacchetta; Anya Tsalenko; Douglas Dellinger; Laurakay Bruhn; Matthew H. Porteus

doi:10.1038/nbt.3290

Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells

Ayal Hendel, Rasmus O. Bak, Joseph T. Clark, Andrew B. Kennedy, Daniel E. Ryan, Subhadeep Roy, Israel Steinfeld, Benjamin D. Lunstad, Robert J. Kaiser, Alec B. Wilkens, Rosa Bacchetta, Anya Tsalenko, Douglas Dellinger, Laurakay Bruhn, Matthew H. Porteus

Research output: Contribution to journal › Article › peer-review

Abstract

CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facilitated by the Cas endonuclease. Here we report that chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in human primary T cells and CD34 + hematopoietic stem and progenitor cells. Co-delivering chemically modified sgRNAs with Cas9 mRNA or protein is an efficient RNA-or ribonucleoprotein (RNP)-based delivery method for the CRISPR-Cas system, without the toxicity associated with DNA delivery. This approach is a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biotechnological and therapeutic applications of the CRISPR-Cas technology.

Original language	English
Pages (from-to)	985-989
Number of pages	5
Journal	Nature biotechnology
Volume	33
Issue number	9
DOIs	https://doi.org/10.1038/nbt.3290
State	Published - 10 Sep 2015
Externally published	Yes

All Science Journal Classification (ASJC) codes

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology
Molecular Medicine
Biomedical Engineering

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10.1038/nbt.3290

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Hendel, A., Bak, R. O., Clark, J. T., Kennedy, A. B., Ryan, D. E., Roy, S., Steinfeld, I., Lunstad, B. D., Kaiser, R. J., Wilkens, A. B., Bacchetta, R., Tsalenko, A., Dellinger, D., Bruhn, L., & Porteus, M. H. (2015). Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nature biotechnology, 33(9), 985-989. https://doi.org/10.1038/nbt.3290

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title = "Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells",

abstract = "CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facilitated by the Cas endonuclease. Here we report that chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in human primary T cells and CD34 + hematopoietic stem and progenitor cells. Co-delivering chemically modified sgRNAs with Cas9 mRNA or protein is an efficient RNA-or ribonucleoprotein (RNP)-based delivery method for the CRISPR-Cas system, without the toxicity associated with DNA delivery. This approach is a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biotechnological and therapeutic applications of the CRISPR-Cas technology.",

author = "Ayal Hendel and Bak, {Rasmus O.} and Clark, {Joseph T.} and Kennedy, {Andrew B.} and Ryan, {Daniel E.} and Subhadeep Roy and Israel Steinfeld and Lunstad, {Benjamin D.} and Kaiser, {Robert J.} and Wilkens, {Alec B.} and Rosa Bacchetta and Anya Tsalenko and Douglas Dellinger and Laurakay Bruhn and Porteus, {Matthew H.}",

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doi = "10.1038/nbt.3290",

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AU - Hendel, Ayal

AU - Bak, Rasmus O.

AU - Clark, Joseph T.

AU - Kennedy, Andrew B.

AU - Ryan, Daniel E.

AU - Roy, Subhadeep

AU - Steinfeld, Israel

AU - Lunstad, Benjamin D.

AU - Kaiser, Robert J.

AU - Wilkens, Alec B.

AU - Bacchetta, Rosa

AU - Tsalenko, Anya

AU - Dellinger, Douglas

AU - Bruhn, Laurakay

AU - Porteus, Matthew H.

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AB - CRISPR-Cas-mediated genome editing relies on guide RNAs that direct site-specific DNA cleavage facilitated by the Cas endonuclease. Here we report that chemical alterations to synthesized single guide RNAs (sgRNAs) enhance genome editing efficiency in human primary T cells and CD34 + hematopoietic stem and progenitor cells. Co-delivering chemically modified sgRNAs with Cas9 mRNA or protein is an efficient RNA-or ribonucleoprotein (RNP)-based delivery method for the CRISPR-Cas system, without the toxicity associated with DNA delivery. This approach is a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biotechnological and therapeutic applications of the CRISPR-Cas technology.

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JO - Nature biotechnology

JF - Nature biotechnology

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Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells

Abstract

All Science Journal Classification (ASJC) codes

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