CRISPR-mediated live imaging of genome editing and transcription-Reference-Cited by-同舟云学术

CRISPR-mediated live imaging of genome editing and transcription

Published:2019-09-20 Issue:6459 Volume:365 Page:1301-1305
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Wang Haifeng¹^ORCID,Nakamura Muneaki¹,Abbott Timothy R.¹^ORCID,Zhao Dehua¹,Luo Kaiwen¹²^ORCID,Yu Cordelia¹³,Nguyen Cindy M.¹^ORCID,Lo Albert¹^ORCID,Daley Timothy P.¹⁴^ORCID,La Russa Marie¹,Liu Yanxia¹^ORCID,Qi Lei S.¹⁵⁶^ORCID

Affiliation:

1. Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.

2. ZJU-UOE Institute, Zhejiang University, School of Medicine, Haining, China.

3. Castilleja School, Palo Alto, CA 94301, USA.

4. Department of Statistics, Stanford University, Stanford, CA 94305, USA.

5. Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305, USA.

6. ChEM-H Institute, Stanford University, Stanford, CA 94305, USA.

Abstract

Tracking nucleic acids in living cells Fluorescence in situ hybridization (FISH) is a powerful molecular technique for detecting nucleic acids in cells. However, it requires cell fixation and denaturation. Wang et al. found that CRISPR-Cas9 protects guide RNAs from degradation in cells only when bound to target DNA. Taking advantage of this target-dependent stability switch, they developed a labeling technique, named CRISPR LiveFISH, to detect DNA and RNA using fluorophore-conjugated guide RNAs with Cas9 and Cas13, respectively. CRISPR LiveFISH improves the signal-to-noise ratio, is compatible with living cells, and allows tracking real-time dynamics of genome editing, chromosome translocation, and transcription. Science , this issue p. 1301

Funder

NIH Office of the Director

Pew Charitable Trusts

Alfred P. Sloan Foundation

Li Ka Shing Foundation

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

Reference31 articles.

1. Targeted genomic rearrangements using CRISPR/Cas technology

2. Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements