title: The CRISPR system--keeping zebrafish gene targeting fresh.
abstract: We are entering a new era in our ability to modify and edit the genomes of model organisms. Zinc finger nucleases (ZFNs) opened the door to the first custom nuclease-targeted genome engineering in the late 1990s. However, ZFNs remained out of reach for most research labs because of the difficulty of production, high costs, and modest efficacy in many applications. Transcription activator-like effector nucleases (TALENs) were built upon a DNA binding system discovered in a group of plant bacterial pathogens and broadened custom nuclease technology, showing significant improvements in both targeting flexibility and efficiency. Perhaps most importantly, TALENs are open source and easy to produce, providing zebrafish laboratories around the world with affordable tools that can be made in-house rapidly, at low cost, and with reliably high activity. Now a new system for targeted genome engineering derived from the CRISPR/Cas system in eubacteria and archaea promises to simplify this process further. Together, these tools will help overcome many of the bottlenecks that have constrained gene targeting in zebrafish, paving the way for advanced genome engineering applications in this model teleost.
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Addiction Research Center, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Blackburn
authors.forename: Jarryd M
authors.initials: JM
authors.lastname: Campbell
authors.forename: Karl J
authors.initials: KJ
authors.lastname: Clark
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Ekker
mesh_terms: D000818:Animals; D018390:Gene Targeting / Q000379:methods*; D016678:Genome; D055029:Inverted Repeat Sequences; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism
references.citation: Nat Methods. 2013 Apr;10(4):329-31
references.pmid: 23435258
references.citation: Nature. 2012 Nov 1;491(7422):114-8
references.pmid: 23000899
references.citation: Nucleic Acids Res. 2011 Jul;39(12):e82
references.pmid: 21493687
references.citation: Nat Biotechnol. 2012 May;30(5):460-5
references.pmid: 22484455
references.citation: Nature. 2012 Feb 15;482(7385):331-8
references.pmid: 22337052
references.citation: Nucleic Acids Res. 2012 Jul;40(12):5560-8
references.pmid: 22373919
references.citation: BMC Bioinformatics. 2013 Jan 16;14:1
references.pmid: 23323762
references.citation: Nat Methods. 2012 Jun;9(6):588-90
references.pmid: 22543349
references.citation: PLoS One. 2012;7(5):e37877
references.pmid: 22655075
references.citation: Nucleic Acids Res. 2013 Feb 1;41(4):2769-78
references.pmid: 23303782
references.citation: Science. 2011 Sep 30;333(6051):1843-6
references.pmid: 21960622
references.citation: Science. 2012 Aug 17;337(6096):816-21
references.pmid: 22745249
references.citation: Nucleic Acids Res. 2012 Sep;40(16):8001-10
references.pmid: 22684503
references.citation: Nat Biotechnol. 2011 Aug 05;29(8):697-8
references.pmid: 21822241
references.citation: Zebrafish. 2013 Mar;10(1):15-20
references.pmid: 23590399
references.citation: Nat Biotechnol. 2013 Mar;31(3):227-9
references.pmid: 23360964
references.citation: Nat Biotechnol. 2011 Aug 05;29(8):699-700
references.pmid: 21822242
references.citation: Cell Stem Cell. 2013 Feb 7;12(2):238-51
references.pmid: 23246482
references.citation: Nat Methods. 2011 Jan;8(1):67-9
references.pmid: 21151135
references.citation: Science. 2013 Feb 15;339(6121):823-6
references.pmid: 23287722
references.citation: Nature. 2011 Mar 31;471(7340):602-7
references.pmid: 21455174