Dynamics of CRISPR-Cas9 genome interrogation in living cells-Reference-Cited by-同舟云学术

Dynamics of CRISPR-Cas9 genome interrogation in living cells

Published:2015-11-13 Issue:6262 Volume:350 Page:823-826
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Knight Spencer C.¹,Xie Liangqi²,Deng Wulan³⁴,Guglielmi Benjamin²,Witkowsky Lea B.²,Bosanac Lana²,Zhang Elisa T.²,El Beheiry Mohamed⁵,Masson Jean-Baptiste³,Dahan Maxime⁴⁵,Liu Zhe³⁴,Doudna Jennifer A.¹²⁶⁷⁸,Tjian Robert²³⁴⁶⁹

Affiliation:

1. Department of Chemistry, University of California, Berkeley, CA, USA.

2. Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.

3. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.

4. Transcriptional Imaging Consortium, Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.

5. Laboratoire Physico-Chimie Curie, Institut Curie, Centre National de la Recherche Scientifique UMR 168, Paris, France.

6. Howard Hughes Medical Institute, Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.

7. Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

8. Innovative Genomics Initiative, University of California, Berkeley, CA, USA.

9. Li Ka Shing Biomedical and Health Sciences Center, University of California, Berkeley, CA, USA.

Abstract

Genome editing with a Cas9 scalpel The Cas9 nuclease forms the heart of the CRISPR-Cas genome editing system. Cas9 binds small guide RNAs that direct it to its target sites, where the nuclease either cleaves or binds to genomic DNA. Knight et al. used single-molecule imaging to track Cas9 in living cells. Cas9 searches the genome for its target sites using rapid threedimensional diffusion. It spends very little time binding to off-target sites, which explains the high accuracy of the CRISPRCas9 editing machine. Science , this issue p. 823

Funder

National Science Foundation

Howard Hughes Medical Institute

California Institute for Regenerative Medicine

Gordon and Betty Moore Foundation

Helen Hay Whitney Foundation

Caribou Biosciences, Inc.

Editas Medicine

Intellia Therapeutics

Publisher

American Association for the Advancement of Science (AAAS)

Subject