title: The new CRISPR-Cas system: RNA-guided genome engineering to efficiently produce any desired genetic alteration in animals.
abstract: The CRISPR-Cas system is the newest targeted nuclease for genome engineering. In less than 1 year, the ease, robustness and efficiency of this method have facilitated an immense range of genetic modifications in most model organisms. Full and conditional gene knock-outs, knock-ins, large chromosomal deletions and subtle mutations can be obtained using combinations of clustered regularly interspaced short palindromic repeats (CRISPRs) and DNA donors. In addition, with CRISPR-Cas compounds, multiple genetic modifications can be introduced seamlessly in a single step. CRISPR-Cas not only brings genome engineering capacities to species such as rodents and livestock in which the existing toolbox was already large, but has also enabled precise genetic engineering of organisms with difficult-to-edit genomes such as zebrafish, and of technically challenging species such as non-human primates. The CRISPR-Cas system allows generation of targeted mutations in mice, even in laboratories with limited or no access to the complex, time-consuming standard technology using mouse embryonic stem cells. Here we summarize the distinct applications of CRISPR-Cas technology for obtaining a variety of genetic modifications in different model organisms, underlining their advantages and limitations relative to other genome editing nucleases. We will guide the reader through the many publications that have seen the light in the first year of CRISPR-Cas technology.
authors.affiliation: Department of Molecular and Cellular Biology, National Centre for Biotechnology (CNB-CSIC), Campus de Cantoblanco, 28049, Madrid, Spain.
authors.forename: Davide
authors.initials: D
authors.lastname: Seruggia
authors.forename: Lluis
authors.initials: L
authors.lastname: Montoliu
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D003851:Deoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D051379:Mice; D011323:Primates; D013045:Species Specificity; D015027:Zebrafish
chemical_list: D003851:Deoxyribonucleases
references.citation: Development. 2013 Dec;140(24):4982-7
references.pmid: 24257628
references.citation: Mol Microbiol. 2002 Mar;43(6):1565-75
references.pmid: 11952905
references.citation: Nature. 2012 Nov 1;491(7422):114-8
references.pmid: 23000899
references.citation: Nat Biotechnol. 2013 Sep;31(9):827-32
references.pmid: 23873081
references.citation: Proc Natl Acad Sci U S A. 2013 Oct 8;110(41):16526-31
references.pmid: 24014591
references.citation: Cell. 2014 Feb 27;156(5):935-49
references.pmid: 24529477
references.citation: Nucleic Acids Res. 2014 Feb;42(4):2577-90
references.pmid: 24270795
references.citation: Nat Biotechnol. 2013 Sep;31(9):822-6
references.pmid: 23792628
references.citation: Cell Stem Cell. 2014 Mar 6;14(3):404-14
references.pmid: 24360884
references.citation: Nature. 2005 Jun 2;435(7042):646-51
references.pmid: 15806097
references.citation: Nat Biotechnol. 2013 Sep;31(9):839-43
references.pmid: 23934178
references.citation: Pigment Cell Res. 2002 Aug;15(4):258-64
references.pmid: 12100491
references.citation: Genes Dev. 1999 Jan 15;13(2):142-5
references.pmid: 9925637
references.citation: Microbiology (Reading). 2005 Aug;151(Pt 8):2551-2561
references.pmid: 16079334
references.citation: Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8929-33
references.pmid: 8799130
references.citation: Biochem Biophys Res Commun. 2014 Mar 21;445(4):791-4
references.pmid: 24491566
references.citation: Nucleic Acids Res. 2011 Jul;39(12):e82
references.pmid: 21493687
references.citation: Cell. 2013 May 9;153(4):910-8
references.pmid: 23643243
references.citation: Science. 2010 Sep 10;329(5997):1355-8
references.pmid: 20829488
references.citation: Curr Biol. 2001 Jun 26;11(12):R455-7
references.pmid: 11448783
references.citation: Proc Natl Acad Sci U S A. 2012 Sep 25;109(39):E2579-86
references.pmid: 22949671
references.citation: Transgenic Res. 1998 Mar;7(2):135-40
references.pmid: 9608741
references.citation: Science. 2008 Dec 19;322(5909):1843-5
references.pmid: 19095942
references.citation: Nucleic Acids Res. 2012 Jul;40(Web Server issue):W117-22
references.pmid: 22693217
references.citation: Nature. 2013 Mar 7;495(7439):50-1
references.pmid: 23467164
references.citation: Proc Natl Acad Sci U S A. 2013 Aug 20;110(34):13904-9
references.pmid: 23918387
references.citation: Science. 2009 Dec 11;326(5959):1501
references.pmid: 19933106
references.citation: PeerJ. 2013 Dec 23;1:e230
references.pmid: 24432195
references.citation: Sci Rep. 2014 Mar 28;4:4513
references.pmid: 24675426
references.citation: Cell Res. 2013 May;23(5):720-3
references.pmid: 23545779
references.citation: Nature. 2011 Jun 15;474(7351):337-42
references.pmid: 21677750
references.citation: Nat Rev Mol Cell Biol. 2013 Jan;14(1):49-55
references.pmid: 23169466
references.citation: Nat Biotechnol. 2014 Jul;32(7):670-6
references.pmid: 24752079
references.citation: Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6064-8
references.pmid: 8016116
references.citation: Nucleic Acids Res. 2013 Aug;41(14):e141
references.pmid: 23748566
references.citation: PLoS One. 2013 Jul 09;8(7):e68708
references.pmid: 23874735
references.citation: Science. 2014 Mar 14;343(6176):1247997
references.pmid: 24505130
references.citation: RNA Biol. 2013 May;10(5):726-37
references.pmid: 23563642
references.citation: J Vis Exp. 2014 Apr 02;(86):
references.pmid: 24747757
references.citation: Nature. 2012 Oct 18;490(7420):407-11
references.pmid: 23023130
references.citation: Cell Stem Cell. 2013 Dec 5;13(6):659-62
references.pmid: 24315440
references.citation: PLoS One. 2014 Mar 05;9(3):e89413
references.pmid: 24598943
references.citation: Cell. 2013 Sep 12;154(6):1370-9
references.pmid: 23992847
references.citation: Cell Res. 2014 Jan;24(1):122-5
references.pmid: 24296780
references.citation: Mol Cell. 2008 Jul 25;31(2):294-301
references.pmid: 18657511
references.citation: Methods Cell Biol. 2011;104:121-7
references.pmid: 21924159
references.citation: Cell. 2014 Feb 13;156(4):836-43
references.pmid: 24486104
references.citation: Nucleic Acids Res. 2013 Oct;41(19):9049-61
references.pmid: 23907390
references.citation: Int J Biochem Cell Biol. 2014 Jan;46:49-55
references.pmid: 24269190
references.citation: PLoS Comput Biol. 2005 Nov;1(6):e60
references.pmid: 16292354
references.citation: Nat Biotechnol. 2014 Mar;32(3):279-284
references.pmid: 24463574
references.citation: PLoS One. 2009;4(5):e5553
references.pmid: 19436741
references.citation: Transgenic Res. 2013 Oct;22(5):965-72
references.pmid: 23494546
references.citation: Genome Res. 2013 Mar;23(3):539-46
references.pmid: 23152450
references.citation: Science. 2012 Aug 17;337(6096):816-21
references.pmid: 22745249
references.citation: Nature. 2010 Nov 4;468(7320):67-71
references.pmid: 21048762
references.citation: Nat Methods. 2014 Apr;11(4):399-402
references.pmid: 24584192
references.citation: PLoS One. 2012;7(9):e41796
references.pmid: 22970113
references.citation: Nature. 2014 Jan 16;505(7483):272
references.pmid: 24429608
references.citation: Nucleic Acids Res. 2013 Nov;41(20):e187
references.pmid: 23997119
references.citation: Genome Res. 2014 Jan;24(1):142-53
references.pmid: 24179142
references.citation: Nat Protoc. 2013 Nov;8(11):2281-2308
references.pmid: 24157548
references.citation: Cell Stem Cell. 2013 Apr 4;12(4):393-4
references.pmid: 23561441
references.citation: Nat Biotechnol. 2013 Sep;31(9):833-8
references.pmid: 23907171
references.citation: Nat Biotechnol. 2013 Aug;31(8):681-3
references.pmid: 23929336
references.citation: Nat Biotechnol. 2008 Jun;26(6):695-701
references.pmid: 18500337
references.citation: Transgenic Res. 2010 Jun;19(3):363-71
references.pmid: 19821047
references.citation: J Bacteriol. 1987 Dec;169(12):5429-33
references.pmid: 3316184
references.citation: Nat Rev Microbiol. 2011 Jun;9(6):467-77
references.pmid: 21552286
references.citation: Science. 2013 Feb 15;339(6121):819-23
references.pmid: 23287718
references.citation: Cell. 2013 Sep 12;154(6):1380-9
references.pmid: 23992846
references.citation: Nat Rev Mol Cell Biol. 2003 Sep;4(9):712-20
references.pmid: 14506474