Home LiteratureArticle Details
PMID: 23360964 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Efficient genome editing in zebrafish using a CRISPR-Cas system.

Nature biotechnology ·Vol. 31 ·No. 3 ·2013-03-00 ·Pages 227-9

Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT, Yeh JR, Joung JK

Abstract

In bacteria, foreign nucleic acids are silenced by clustered, regularly interspaced, short palindromic repeats (CRISPR)--CRISPR-associated (Cas) systems. Bacterial type II CRISPR systems have been adapted to create guide RNAs that direct site-specific DNA cleavage by the Cas9 endonuclease in cultured cells. Here we show that the CRISPR-Cas system functions in vivo to induce targeted genetic modifications in zebrafish embryos with efficiencies similar to those obtained using zinc finger nucleases and transcription activator-like effector nucleases.

MeSH Terms
Animals Base Sequence CRISPR-Cas Systems DNA/genetics DNA Cleavage Embryo, Nonmammalian Endonucleases/genetics Genetic Engineering Genome Inverted Repeat Sequences Molecular Sequence Data Mutation Nucleic Acid Conformation RNA Editing RNA, Guide/genetics Zebrafish/genetics
Chemicals
RNA, Guide DNA Endonucleases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Hwang Woong Y
Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts, USA.
Fu Yanfang
Reyon Deepak
Maeder Morgan L
Tsai Shengdar Q
Sander Jeffry D
Peterson Randall T
Yeh J-R Joanna
Joung J Keith
References (21)
21 references, click to expand
  1. CRISPR-based adaptive immune systems.
    Curr Opin Microbiol. 2011 Jun;14(3):321-7 PMID: 21531607
  2. Heritable gene targeting in zebrafish using customized TALENs.
    Nat Biotechnol. 2011 Aug 05;29(8):699-700 PMID: 21822242
  3. Genome editing with engineered zinc finger nucleases.
    Nat Rev Genet. 2010 Sep;11(9):636-46 PMID: 20717154
  4. FLASH assembly of TALENs for high-throughput genome editing.
    Nat Biotechnol. 2012 May;30(5):460-5 PMID: 22484455
  5. CRISPR provides acquired resistance against viruses in prokaryotes.
    Science. 2007 Mar 23;315(5819):1709-12 PMID: 17379808
  6. Mfold web server for nucleic acid folding and hybridization prediction.
    Nucleic Acids Res. 2003 Jul 1;31(13):3406-15 PMID: 12824337
  7. ZiFiT (Zinc Finger Targeter): an updated zinc finger engineering tool.
    Nucleic Acids Res. 2010 Jul;38(Web Server issue):W462-8 PMID: 20435679
  8. Small CRISPR RNAs guide antiviral defense in prokaryotes.
    Science. 2008 Aug 15;321(5891):960-4 PMID: 18703739
  9. Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA).
    Nat Methods. 2011 Jan;8(1):67-9 PMID: 21151135
  10. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
    Science. 2012 Aug 17;337(6096):816-21 PMID: 22745249
  11. Highly efficient generation of heritable zebrafish gene mutations using homo- and heterodimeric TALENs.
    Nucleic Acids Res. 2012 Sep;40(16):8001-10 PMID: 22684503
  12. Targeted gene disruption in somatic zebrafish cells using engineered TALENs.
    Nat Biotechnol. 2011 Aug 05;29(8):697-8 PMID: 21822241
  13. Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria.
    Proc Natl Acad Sci U S A. 2012 Sep 25;109(39):E2579-86 PMID: 22949671
  14. RNA-guided genetic silencing systems in bacteria and archaea.
    Nature. 2012 Feb 15;482(7385):331-8 PMID: 22337052
  15. Excess information at bacteriophage T7 genomic promoters detected by a random cloning technique.
    Nucleic Acids Res. 1989 Jan 25;17(2):659-74 PMID: 2915926
  16. Targeted mutagenesis in zebrafish using customized zinc-finger nucleases.
    Nat Protoc. 2009;4(12):1855-67 PMID: 20010934
  17. Simple methods for generating and detecting locus-specific mutations induced with TALENs in the zebrafish genome.
    PLoS Genet. 2012;8(8):e1002861 PMID: 22916025
  18. CRISPR/Cas, the immune system of bacteria and archaea.
    Science. 2010 Jan 8;327(5962):167-70 PMID: 20056882
  19. TALENs: a widely applicable technology for targeted genome editing.
    Nat Rev Mol Cell Biol. 2013 Jan;14(1):49-55 PMID: 23169466
  20. Zinc Finger Targeter (ZiFiT): an engineered zinc finger/target site design tool.
    Nucleic Acids Res. 2007 Jul;35(Web Server issue):W599-605 PMID: 17526515
  21. Rapid mutation of endogenous zebrafish genes using zinc finger nucleases made by Oligomerized Pool ENgineering (OPEN).
    PLoS One. 2009;4(2):e4348 PMID: 19198653
Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1546-1696
DOI
10.1038/nbt.2501
Published
2013-03-00
Epub
2013-00-29
Pages
227-9
Language
English
Region
United States
NLM ID
9604648
PMCID
PMC3686313
Subset
IM
Grants
NIGMS NIH HHS · DP1 GM105378 · United States
NHGRI NIH HHS · P50 HG005550 · United States
NCI NIH HHS · R01 CA140188 · United States
NIA NIH HHS · K01 AG031300 · United States
NCCDPHP CDC HHS · DP1 GM105378 · United States
NIGMS NIH HHS · R01 GM088040 · United States
NIGMS NIH HHS · F32 GM105189 · United States
Corrections
CommentIn
PMID: 23565557
External Links
PubMed source DOI: 10.1038/nbt.2501 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2