B6;SJL-Prlh^em2Mgmj/J

Stock number: 036748
Product name: Prlh^Flox
Research areas: Cancer Research, Neurobiology Research, Research Tools

Description

Prlh^Flox mice have loxP sites flanking exon 3 of the prolactin releasing hormone (Prlh) gene. Exposure to Cre recombinase removes the floxed sequence - creating a null allele. These mice may be useful for studying feeding behavior, obesity, and metabolism.

Detailed description

The Prlh gene encodes encodes the anorectic prolactin releasing peptide (PRRP), which is involved in regulation of metabolism and feeding behavior.

To create the Prlh^Flox allele, CRISPR/Cas9 endonuclease-mediated genome editing of Prlh was used to introduce loxP sites flanking exon 3. Prior to Cre recombinase exposure, mice heterozygous or homozygous for this floxed allele are viable and fertile, with no reported phenotypic abnormalities. No alterations of endogenous gene expression were observed.

Upon exposure to Cre recombinase, deletion of the floxed sequences results in a null allele.
For example, according to the donating investigator, when Prlh^Flox mice were bred to Calcr^Cre-expressing mice (Stock No. 037028), the resulting Prlh^CalcrKO knock-out mice exhibited a ~90% reduction in PRRP expression in the nucleus of the solitary tract. Those Prlh^CalcrKO mice show increased body weights when fed high fat diet (60 kcal% fat), but show no difference in total chow intake and no difference in body weight when fed standard rodent chow (Cheng et al., 2021).

Development

The Prlh^Flox knock-in allele was created in the laboratory of Dr. Martin Myers (University of Michigan) to have loxP sites flanking the second coding exon of the prolactin releasing hormone locus (Prlh) on chromosome 1. The specific details are below.

The Prlh^Flox allele was generated using CRISPR/Cas9 endonuclease-mediated genome editing. Guide RNAs were selected to target the second coding exon of the Prlh gene. Double-stranded oligonucleotide donor DNAs were created containing loxP sequences. These sequences and Cas9 protein were introduced into single cell B6SJLF2 zygotes and transferred to pseudopregnant females. Progeny were screened by DNA sequencing to identify correctly targeted pups, which were then bred to C57BL/6J mice (Stock No. 000664) for germline transmission. The Prlh^Flox colony was backcrossed to C57BL/6J for a total of at least 2 generations prior to sending males to The Jackson Laboratory Repository in 2022. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Prlh^em2Mgmj
Name: endonuclease-mediated mutation 2, Martin G Myers, Jr
MGI accession ID: MGI:6757219
Generation method: Endonuclease-mediated
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Prlh^Flox
Marker symbol: Prlh
Marker name: prolactin releasing hormone
Marker synonyms: EG623503; PRH; prrp
Chromosome: 1
Strain of origin: (C57BL/6 x SJL)F2
Molecular note: CRISPR/cas9 genome editing is used to insert LoxP sites upstream and downstream of Prlh exon 2.