Recombination between inverted loxP sites is cytotoxic for proliferating cells and provides a simple tool for conditional cell ablation.
The loxP/Cre recombination system is a widely used tool for mouse functional genomics, in particular for in vivo conditional mutagenesis. Depending on the relative orientation and position of loxP sites, Cre-mediated recombination can result in a variety of targeted genomic rearrangements. It was previously reported that loss of the loxP-carrying chromosome can occur when loxP sites are arranged in inverse orientation. By using a chromosome 2 carrying inverted loxP sites, we found that Cre-mediated recombination not only causes chromosomal loss but also triggers apoptosis. We show that targeted recombination between inverted loxP sites (TRIP) triggers cell death specifically in proliferating Cre-expressing cells, and we provide evidence that TRIP is an efficient tool to ablate proliferating cells within genetically defined cell populations. Furthermore, the procedure requires only a simple, one-step intercross but neither the use of toxins nor the additional step of prodrug injection. With the large repertoire of tissue-specific or inducible Cre-expressing transgenes available, TRIP-mediated cell ablation is valuable to investigate the function of a large variety of cell populations in the context of a whole organism, which includes mechanisms underlying organ development and tissue homeostasis.
Grégoire
Damien
D
Laboratory of Genetics and Development, Institut de Recherches Cliniques de Montréal, 110 Avenue des Pins Ouest, H2W1R7, Université de Montréal, Montréal Québec, Canada.
Kmita
Marie
M
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics; D002872:Chromosome Deletion; D004622:Embryo, Mammalian / Q000166:cytology; D019426:Integrases / Q000378:metabolism*; D051379:Mice; D009006:Monosomy / Q000235:genetics*; D009928:Organ Specificity; D011995:Recombination, Genetic
C045073:Cre recombinase; D019426:Integrases
Nature. 2002 Nov 14;420(6912):145-50
12432383
Nat Genet. 1997 Oct;17(2):223-5
9326948
Nat Rev Genet. 2001 Oct;2(10):780-90
11584294
Mol Vis. 2006 Apr 18;12:389-98
16636658
Anat Rec. 1985 Jun;212(2):199-205
3842042
Science. 1987 Dec 11;238(4833):1563-5
3685993
Dev Cell. 2004 Jan;6(1):7-28
14723844
J Cell Biol. 1998 Oct 5;143(1):253-66
9763436
Curr Protoc Hum Genet. 2001 May;Chapter 4:Unit4.10
18428279
Curr Protoc Cell Biol. 2004 Sep;Chapter 22:Unit 22.4
18228455
Nat Genet. 1998 Dec;20(4):381-4
9843213
Genesis. 2002 Jun;33(2):77-80
12112875
Nat Methods. 2005 Jun;2(6):419-26
15908920
Curr Biol. 1998 Dec 3;8(24):1323-6
9843687
J Biochem. 2006 Dec;140(6):831-41
17071947
Genesis. 2000 Nov-Dec;28(3-4):147-55
11105057
Nat Biotechnol. 2001 Aug;19(8):746-50
11479567
J Exp Zool. 1985 Dec;236(3):353-60
4086989
Cell Cycle. 2004 Sep;3(9):1104-6
15326371
Genesis. 2005 Nov;43(3):129-35
16267821
Nat Methods. 2007 Jan;4(1):23-5
17086180
Cell. 1987 Jul 31;50(3):435-43
3649277
Nat Genet. 2000 Dec;26(4):451-4
11101844