B6(Cg)-Parp1^{tm1c(EUCOMM)Hmgu}/WlkrJ

Stock number: 032650
Product name: Parp1^loxP
Research areas: Apoptosis Research, Cancer Research, Cell Biology Research, Diabetes and Obesity Research, Immunology, Inflammation and Autoimmunity Research, Neurobiology Research, Research Tools

Description

Parp1^loxP is a floxed allele with loxP sites flanking exon 4 of the poly (ADP-ribose) polymerase family, member 1 locus. Cre recombinase-mediated removal of the floxed sequence creates a PARP-1 knock-out allele. Parp1^loxP mice may be useful in studying poly(ADP-ribosyl)ation (PARylation) in a variety of nuclear processes (including transcription, RNA processing and DNA repair).

Detailed description

Poly(ADP-ribosyl)ation (PARylation) is the post-translational modification of proteins mediated by PARP family members. Parp1 encodes poly(ADP-ribose) polymerase-1 (PARP-1 or ARTD1), an abundant nuclear protein involved in a variety of nuclear processes, including transcription, RNA processing and DNA repair.

The Parp1^loxP allele has loxP sites flanking Parp1 exon 4. Mice heterozygous or homozygous for this floxed allele are viable and fertile with normal breeding, and no reported gross physical or behavioral abnormalities.

Upon exposure to Cre recombinase, the floxed region is deleted, creating a knock-out allele. For example, when Parp1^loxP are bred to Pdgfra-cre/ERT transgenic mice (Stock No. 018280), the resulting Parp1^loxP/loxP;Pdgfra-cre/ERT mice have the tamoxifen-inducible deletion of PARP-1 in primary preadipocytes isolated from the stromal vascular fraction of adipose tissue. This allows studies of the specific role of PARP-1 in adipogenesis, separated from any potential complicating developmental effects of global PARP-1 deletion. [Luo et al. 2017 Mol Cell 65:260]

Development

Mice harboring the Parp1^loxP allele (floxed exon 4 ; Parp1^{tm1c(EUCOMM)Hmg}) were created in the laboratory of Dr. W. Lee Kraus (University of Texas Southwestern Medical Center). First, they obtained C57BL/6N-derived embryonic stem (ES) cells with the Parp1^{tm1a(EUCOMM)Hmg} targeted mutation designed to have a frt-SA-lacZ-pA-loxP-neo-pA-loxP sequence upstream of exon 4, and a loxP site downstream of exon 4 of the poly (ADP-ribose) polymerase family, member 1 locus (Parp1) on chromosome 1. Dr. Kraus' laboratory generated chimeric animals and bred them to C57BL/6N mice for germline transmission of the Parp1^{tm1a(EUCOMM)Hmg} allele. The donating investigator reports the mice were then bred to C57BL/6J-congenic Rosa26^FLPo mice (obtained from Stock No. 012930) for germline deletion of the frt-flanked sequences. The resulting Parp1^loxP (floxed exon 4 ; Parp1^{tm1c(EUCOMM)Hmg}) colony was then maintained by breeding Parp1^loxP mice together for several generations (and the FLPo-expressing allele was removed). The donating investigator reports the Parp1^loxP colony may have also been bred with Pdgfra-cre/ERT mice (on C57BL/6N genetic background; Stock No. 018280) or LysM-Cre mice (on a mixed C57BL/6J;C57BL/6N genetic background; Stock No. 004781), but their Parp1^loxP colony in 2018 did not retain any cre-expressing alleles/transgenes. In 2018, Parp1^loxP males with black coat color were sent to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6N oocytes (Stock No. 005304).

Allele

Symbol: Parp1^{tm1c(EUCOMM)Hmgu}
Name: targeted mutation 1c, Helmholtz Zentrum Muenchen GmbH
MGI accession ID: MGI:6100929
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed)
Synonyms: Parp1^flox
Marker symbol: Parp1
Marker name: poly (ADP-ribose) polymerase family, member 1
Marker synonyms: 5830444G22Rik; Adprp; ADPRT; ADPRT 1; Adprt1; ARTD1; pADPRT-1; PARP; PARP-1; PARS; Poly-PARP; PPOL; sPARP-1
Chromosome: 1
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 180406272 of Chromosome 1 upstream of the critical exon(s) (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon(s) at position 180407213. The critical exon(s) is/are thus flanked by loxP sites. A "conditional ready" (floxed) allele was created by flp recombinase expression in mice carrying this allele to remove the lacZ sequence and neo selection cassette, leaving loxP sites flanking the critical exon(s).