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 journalArticle

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 languageEnglish
Pages (from-to)985-989
Number of pages5
JournalNature Biotechnology
Volume33
Issue number9
DOIs
Publication statusPublished - Sep 10 2015

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Guide RNA
RNA
Genes
Hematopoietic Stem Cells
CRISPR-Cas Systems
DNA Cleavage
DNA
Ribonucleoproteins
T-cells
Endonucleases
Toxicity
Technology
T-Lymphocytes
Messenger RNA
Proteins
Gene Editing
Therapeutics

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Biotechnology
  • Molecular Medicine
  • Bioengineering
  • Biomedical Engineering

Cite this

Hendel, A., Bak, R. O., Clark, J. T., Kennedy, A. B., Ryan, D. E., Roy, S., ... 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

Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. / Hendel, Ayal; Bak, Rasmus O.; Clark, Joseph T.; Kennedy, Andrew B.; Ryan, Daniel E.; Roy, Subhadeep; Steinfeld, Israel; Lunstad, Benjamin D.; Kaiser, Robert J.; Wilkens, Alec B.; Bacchetta, Rosa; Tsalenko, Anya; Dellinger, Douglas; Bruhn, Laurakay; Porteus, Matthew H.

In: Nature Biotechnology, Vol. 33, No. 9, 10.09.2015, p. 985-989.

Research output: Contribution to journalArticle

Hendel, A, Bak, RO, Clark, JT, Kennedy, AB, Ryan, DE, Roy, S, Steinfeld, I, Lunstad, BD, Kaiser, RJ, Wilkens, AB, Bacchetta, R, Tsalenko, A, Dellinger, D, Bruhn, L & Porteus, MH 2015, 'Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells', Nature Biotechnology, vol. 33, no. 9, pp. 985-989. https://doi.org/10.1038/nbt.3290
Hendel A, Bak RO, Clark JT, Kennedy AB, Ryan DE, Roy S et al. Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. Nature Biotechnology. 2015 Sep 10;33(9):985-989. https://doi.org/10.1038/nbt.3290
Hendel, Ayal ; Bak, Rasmus O. ; Clark, Joseph T. ; Kennedy, Andrew B. ; Ryan, Daniel E. ; Roy, Subhadeep ; Steinfeld, Israel ; Lunstad, Benjamin D. ; Kaiser, Robert J. ; Wilkens, Alec B. ; Bacchetta, Rosa ; Tsalenko, Anya ; Dellinger, Douglas ; Bruhn, Laurakay ; Porteus, Matthew H. / Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells. In: Nature Biotechnology. 2015 ; Vol. 33, No. 9. pp. 985-989.
@article{f3d63011c3714626af38a5a052fc530e,
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.}",
year = "2015",
month = "9",
day = "10",
doi = "10.1038/nbt.3290",
language = "English",
volume = "33",
pages = "985--989",
journal = "Biotechnology",
issn = "1087-0156",
publisher = "Nature Publishing Group",
number = "9",

}

TY - JOUR

T1 - Chemically modified guide RNAs enhance CRISPR-Cas genome editing in human primary cells

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.

PY - 2015/9/10

Y1 - 2015/9/10

N2 - 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.

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.

UR - http://www.scopus.com/inward/record.url?scp=84937905397&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84937905397&partnerID=8YFLogxK

U2 - 10.1038/nbt.3290

DO - 10.1038/nbt.3290

M3 - Article

AN - SCOPUS:84937905397

VL - 33

SP - 985

EP - 989

JO - Biotechnology

JF - Biotechnology

SN - 1087-0156

IS - 9

ER -