Reverse genetics in eukaryotes.
Reverse genetics consists in the modification of the activity of a target gene to analyse the phenotypic consequences. Four main approaches are used towards this goal and will be explained in this review. Two of them are centred on genome alterations. Mutations produced by random chemical or insertional mutagenesis can be screened to recover only mutants in a specific gene of interest. Alternatively, these alterations may be specifically targeted on a gene of interest by HR (homologous recombination). The other two approaches are centred on mRNA. RNA interference is a powerful method to reduce the level of gene products, while MO (morpholino) antisense oligonucleotides alter mRNA metabolism or translation. Some model species, such as Drosophila, are amenable to most of these approaches, whereas other model species are restricted to one of them. For example, in mice and yeasts, gene targeting by HR is prevalent, whereas in Xenopus and zebrafish MO oligonucleotides are mainly used. Genome-wide collections of mutants or inactivated models obtained in several species by these approaches have been made and will help decipher gene functions in the post-genomic era.
Hardy
Serge
S
Université de Rennes 1, Université Européenne de Bretagne, Institut Fédératif de Recherche 140, Rennes, France.
Legagneux
Vincent
V
Audic
Yann
Y
Paillard
Luc
L
D000818:Animals; D030801:Animals, Genetically Modified; D018390:Gene Targeting; D016678:Genome; D051379:Mice; D008957:Models, Genetic; D016376:Oligonucleotides, Antisense / Q000235:genetics; D034622:RNA Interference; D011995:Recombination, Genetic; D015027:Zebrafish
D016376:Oligonucleotides, Antisense
Genome Res. 1999 Sep;9(9):859-67
10508845
Mol Cell Biol. 2007 Feb;27(3):1146-57
17130239
Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2556-61
12589026
Development. 2008 Jan;135(1):159-69
18065431
Hum Mol Genet. 2004 Oct 15;13(20):2409-20
15333583
Plant Cell Physiol. 2010 Apr;51(4):497-523
20208064
Nat Rev Genet. 2006 Jan;7(1):45-54
16369571
Cell. 2009 Feb 20;136(4):642-55
19239886
Methods Mol Biol. 2003;236:205-20
14501067
Nature. 1998 Oct 29;395(6705):854
9804418
Genetics. 2008 Nov;180(3):1275-88
18757930
Nature. 2003 Oct 16;425(6959):686-91
14562095
Nucleic Acids Res. 1999 Nov 15;27(22):4324-7
10536138
Biochem Biophys Res Commun. 2000 Mar 5;269(1):64-9
10694478
Plant Physiol. 2001 Jun;126(2):480-4
11402178
Nucleic Acids Res. 2007;35(2):e11
17164286
Nucleic Acids Res. 2009 Oct;37(19):e126
19671523
EMBO J. 2007 Jan 10;26(1):170-83
17159906
Genes Dev. 2008 Jan 15;22(2):141-51
18198333
Science. 2000 Jun 16;288(5473):2013-8
10856208
Cell. 2007 Jan 12;128(1):9-13
17218247
Genome Res. 2003 Dec;13(12):2700-7
14613981
Nature. 2007 Jul 12;448(7150):151-6
17625558
Methods. 2001 Jul;24(3):218-29
11403571
PLoS One. 2006 Dec 20;1:e104
17218990
Methods. 2006 Jul;39(3):220-7
16828311
Genetics. 1999 Sep;153(1):135-77
10471706
Science. 1998 Oct 16;282(5388):430-1
9841401
Nucleic Acids Res. 2004 Aug 13;32(14):e117
15310838
Nat Methods. 2006 Oct;3(10):839-44
16990816
Cell. 2007 Apr 20;129(2):235
17448981
Nucleic Acids Res. 2006 Jan 1;34(Database issue):D642-8
16381950
Nat Genet. 2006 Aug;38(8):896-903
16845399
Nat Genet. 2004 Nov;36(11):1231-7
15502826
Curr Opin Cell Biol. 2009 Jun;21(3):394-402
19359157
Nature. 2000 Nov 16;408(6810):331-6
11099034
Front Biosci (Landmark Ed). 2009 Jan 01;14:225-36
19273065
PLoS Genet. 2007 May 25;3(5):e78
17530925
Development. 2008 May;135(10):1735-43
18403413
Plant Mol Biol. 2003 Sep;53(1-2):247-59
14756321
Nature. 2003 Oct 16;425(6959):737-41
14562106
Nature. 2001 May 24;411(6836):494-8
11373684
Mol Gen Genet. 1984;197(2):345-6
6394957
RNA. 2010 Jan;16(1):10-5
19933768
Proc Natl Acad Sci U S A. 2010 Jan 5;107(1):157-62
19966286
Nature. 2006 Sep 21;443(7109):359-63
16964239
PLoS Genet. 2006 Nov 17;2(11):e193
17112317
Genome Biol. 2007;8 Suppl 1:S9
18047701
Science. 2003 Aug 1;301(5633):653-7
12893945
Annu Rev Biochem. 2010;79:295-319
20205586
Nucleic Acids Res. 2004 Feb 24;32(4):e40
14982959
Nat Rev Genet. 2009 Feb;10(2):94-108
19148191
Nature. 2002 Jan 10;415(6868):180-3
11805837
Nucleic Acids Res. 2009 Jul;37(13):e93
19483100
PLoS One. 2010 Jan 28;5(1):e8928
20126626
Nature. 2006 Mar 30;440(7084):637-43
16554755
Cell. 2009 May 29;137(5):961-71
19490899
Genetics. 2004 Jun;167(2):761-81
15238527
Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7431-5
8395047
Methods Mol Biol. 2009;561:245-63
19504076
PLoS Biol. 2005 Aug;3(8):e254
16089502
Nat Genet. 2004 Mar;36(3):288-92
14981519
Nat Genet. 2008 Nov;40(11):1375-83
18953339
Genetics. 2004 Nov;168(3):1477-89
15579700
Nature. 2003 Jan 16;421(6920):231-7
12529635
Int J Dev Biol. 2009;53(7):1045-51
19598121
Mech Dev. 2006 Jul;123(7):513-29
16859902
Nature. 2002 Jan 10;415(6868):141-7
11805826
Nat Genet. 2004 Mar;36(3):283-7
14981521
Genes Dev. 2002 Mar 15;16(6):707-19
11914276
Nat Methods. 2010 Jun;7(6):451-3
20418868
Genetics. 1993 Dec;135(4):973-80
8307337
Biochem Biophys Res Commun. 2007 Jun 29;358(2):521-7
17493584
Nat Methods. 2006 Feb;3(2):109-16
16432520
Plant Cell. 1999 Dec;11(12):2283-90
10590158
Nature. 1998 Feb 19;391(6669):806-11
9486653
Genetics. 2009 Nov;183(3):1165-73
19720858
Nature. 2002 Jul 25;418(6896):387-91
12140549
Nat Rev Genet. 2005 Jun;6(6):507-12
15931173
Proc Natl Acad Sci U S A. 2009 Nov 3;106(44):18662-7
19858493
Development. 2004 Dec;131(24):6107-17
15548579
Plant Physiol. 2000 Jun;123(2):439-42
10859174
Science. 2004 Feb 6;303(5659):808-13
14764870
Genome Biol. 2007;8(9):R203
17903264
Genesis. 2006 May;44(5):252-61
16676321
J Biol. 2007;6(3):8
17897480
Nat Genet. 1997 Sep;17(1):119-21
9288111
Handb Exp Pharmacol. 2007;(178):3-28
17203649
PLoS One. 2009;4(3):e4761
19283079
Nat Genet. 2007 Feb;39(2):259-63
17220889
Science. 1999 Aug 6;285(5429):901-6
10436161
BMC Genomics. 2006 Oct 18;7:262
17049087
Annu Rev Genet. 2009;43:601-25
19712041
Genetica. 2009 Sep;137(1):39-46
19343510
Plant Cell. 1990 Apr;2(4):279-289
12354959
Proc Natl Acad Sci U S A. 2004 Aug 31;101(35):12792-7
15256591
Biochim Biophys Acta. 2010 Oct;1802(10):796-807
20004244
BMC Plant Biol. 2009 Dec 14;9:147
20003424
Science. 2007 Oct 12;318(5848):271-4
17761850
Nucleic Acids Res. 2004 Sep 08;32(16):e128
15356288
Fly (Austin). 2008 Nov-Dec;2(6):300-2
19098435
J Biol Chem. 2009 Nov 20;284(47):32370-83
19762469
Mol Cell Biol. 1983 Apr;3(4):747-9
6343844
Zebrafish. 2009 Mar;6(1):69-77
19374550
Genome Res. 2004 Oct;14(10B):2176-89
15489341
PLoS Genet. 2006 Jun;2(6):e91
16789825
Nat Biotechnol. 1999 Oct;17(10):1030-2
10504710
Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929-33
347451
Science. 2008 Oct 17;322(5900):405-10
18818364
Genome Res. 2005 May;15(5):718-23
15867432