B6;DBA-Scrt2^em1Zyliu/J

Stock number: 034390
Product name: Scrt2-tdTomato
Research areas: Neurobiology Research, Research Tools

Description

Scrt2-tdTomato mice have a CRISPR/Cas9-generated P2A-tdtomato cassette inserted immediately before the stop codon of the scratch family zinc finger 2 (Scrt2) gene. These mice are useful for visualizing inner ear cochlear spiral ganglion neurons (SGNs) and as a tool for studying SGN developmental and regenerative studies.

Detailed description

Scrt2-tdTomato mice have a CRISPR/Cas9-generated P2A-tdtomato cassette inserted immediately before the stop codon of the scratch family zinc finger 2 (Scrt2) gene. Insertion of this reporter cassette does not affect endogenous Scrt2 gene expression. Scrt2 encodes a transcriptional repressor involved in brain neurogenesis. In these mice, tdTomato is evident beginning between E10.5 and E13.5 permanently and specifically in inner ear cochlear spiral ganglion neurons (SGNs). tdTomato expression in these mice is visible by immunohistochemistry. Homozygous mice are viable and fertile.

Development

Plasmids encoding a sgRNA (5'-GCCTCGGCGGGCATCCCGCA-3'), designed to introduce a viral P2A oligopeptide (P2A)-tdTomato cassette immediately before the stop codon of the scratch family zinc finger 2 (Scrt2) gene, and the cas9 nuclease were introduced into the cytoplasm of C57BL/6/DBA-derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females and pups were identified by PCR. These Scrt2-tdTomato mice were further bred to C57BL/6NCrl mice to develop the colony. Southern-blot analysis showed that there were no random insertions of donor DNA in the genome. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ (Stock No. 005304) oocytes.

Allele

Symbol: Scrt2^em1Zyliu
Name: endonuclease-mediated mutation 1, Zhiyong Liu
MGI accession ID: MGI:6368164
Generation method: Endonuclease-mediated
Attributes: Reporter
Synonyms: Scrt2-tdTomato
Marker symbol: Scrt2
Marker name: scratch family zinc finger 2
Marker synonyms: RGD1564796; ZNF898B
Chromosome: 2
Strain of origin: C57BL/6 x DBA
Site of expression: Inner ear cochlear spiral ganglion neurons (SGNs).
Molecular note: Using CRISPR/Cas9 technology with sgRNA 5'-GCCTCGGCGGGCATCCCGCA-3', sequence for the P2A self-cleaving peptide sequence plus the tdTomato red fluorescent reporter gene was inserted immediately before the stop codon.