B6;129S6-Pou4f2^tm4(DTA)Whk/J

Stock number: 038952
Product name: brn3b^Z-dta
Research areas: Cell Biology Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

Brn3b^Z-dta conditional knock-in/knock-out mice express a lacZ reporter prior to exposure to Cre recombinase and diphtheria toxin (DTA) post Cre recombination, under the direction of the Pou4f2 (POU domain, class 4, transcription factor 2, also known as Brn3b) promoter, resulting in the ablation of newly formed retinal ganglion cells. LacZ expression recapitulates Pou4f2 expression.

Detailed description

Pou4f2 encodes a transcription factor expressed in the retina. It is one of the earliest genes expressed in retinal ganglion cells immediately after birth.

Brn3b^Z-dta conditional knock-in/knock-out mice express a lacZ reporter prior to exposure to Cre recombinase and diphtheria toxin subunit alpha (DTA) post Cre recombination under the direction of the Pou4f2 (POU domain, class 4, transcription factor 2, also known as Brn3b) promoter. The insertion of the lacZ-DTA cassette replaces the Pour4f2 coding sequence, thus creating a null allele. LacZ expression recapitulates Pou4f2 expression. Mice homozygous for the mutation are viable and fertile, but have defective retinal ganglion cell differentiation. Mice heterozygous for the mutation are normal, but when they are bred to mice that express tissue-specific Cre recombinase in tissues where cre and Pou4f2 expression intersect, resulting offspring will express DTA in the cre-expressing tissues. When combined with mice expressing Cre recombinase in the developing retina (STOCK Tg(Six3-cre)69Frty/GcoJ, (Stock No. 019755), DTA will ablate retinal ganglion cells as they are generated in double mutant mice.

Development

The brn3b^Z-dta targeting construct was designed to replace the Pou4f2 (POU domain, class 4, transcription factor 2, also known as Brn3b) coding region on chromosome 8 with from 5' to 3': a loxP site, lacZ reporter with a stop codon (TAA) at the 3' end, a second loxP site, DTA, and a PGK-neomycin selection cassette. The floxed lacZ reporter is inserted immediately downstream of the dta translation initiation codon (ATG). The construct was electroporated into (129S6/SvEvTac x C57BL/6NCrl)F1-derived G4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to C57BL/6J females. Mice were maintained on a mixed genetic background by the donating lab. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize B6129SF1/J oocytes (Stock No. 101043).

Allele

Symbol: Pou4f2^tm4(DTA)Whk
Name: targeted mutation 4, William H Klein
MGI accession ID: MGI:3838536
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter
Synonyms: brn3b^Z-dta; Pou4f2^lacZ-dta; Pou4f2^tm4Whk
Marker symbol: Pou4f2
Marker name: POU domain, class 4, transcription factor 2
Marker synonyms: BRN3.2; Brn-3.2; Brn3b; Brn-3B; mBrn3-3R; Pou4f-rs1
Chromosome: 8
Strain of origin: 129S/SvEv
Molecular note: The coding sequence was replaced by a floxed lacZ with a stop codon, a diphtheria toxin A gene, and a neomycin selection cassette. As long as lacZ remains in place, the diphtheria toxin A gene is not expressed.