B6(129)-Igs2^{tm1(CAG-AU040320/mCherry,-cas9*)Janc}/J

Stock number: 037553
Product name: SELECTIV
Research areas: Neurobiology Research, Research Tools

Description

SELECTIV (SELective Expression and Controlled Transduction In Vivo) mice conditionally express the adeno-associated virus receptor (AAVR, formally AU040320) fused to the mCherry reporter and spCas9. This strain provides cre-inducible overexpression of AAVR allowing efficient and tissue-specific expression of transgenes using AAV vectors. In addition, SELECTIV mice can be used for testing of AAV vectors for human gene therapy.

Of note, the null allele of Aavr (AU040320) is distributed as Stock No. 037596.

Detailed description

SELECTIV (SELective Expression and Controlled Transduction In Vivo) mice conditionally express the adeno-associated virus receptor (AAVR, formally AU040320) fused to the mCherry reporter as well as a mammalian codon-optimized cas9 gene (derived from Streptococcus pyogenes CRISPR associated protein 9 [spCas9]). Transduction of the AAV (adeno-associated virus) vector for in vivo transduction by most AAV (adeno-associated virus) vectors requires the expression of its receptor, AAVR. The SELECTIV controls AAV transduction through the regulation of its receptor. The presence of spCas9 in the construct allows gene knockout experiments that use AAV delivery of a sgRNA targeted against a gene of interest.

Crossing SELECTIV mice to mice carrying tissue specific Cre recombinase removes the floxed stop (LSL) and allows the specific expression of AAVR in progeny. AAVR-expressing mice are then injected with an AAV vector linked to a gene of interest resulting in transgene expression in cre-expressing tissues. This strain provides cre-inducible overexpression of AAVR allowing efficient and tissue-specific expression of transgenes using AAV vectors. Cells that overexpress AAVR are transduced more efficiently in vitro and in vivo. SELECTIV and cre mice can be crossed to Aavr KO mice (B6.FVB-AU040320^em1Janc/J; Stock No. 037596) to also detarget AAV from non-cre cells, allowing for specific transduction of only cre-expressing cells.

 

Development

The SELECTIV mouse was generated using PhiC31 integrase-mediated transgenesis. A plasmid containing from 5 to 3 a CAG promoter, a loxP-flanked stop sequence (containing a neomycin selection cassette), mouse AU040320 (Aavr) fused to mCherry, an F2A self-cleaving peptide sequence, and a mammalian codon-optimized cas9 gene (derived from Streptococcus pyogenes CRISPR associated protein 9 [spCas9]) and PhiC31 integrase RNA was injected into the cytoplasm of C57BL/6 embryos carrying three tandem attP sites in the "safe harbor" Igs2 (H11) intergenic site (Igs2^tm3.1Luo) on chromosome 11. The resulting progeny were bred to C57BL/6J mice for germline transmission. The colony was maintained on the C57BL/6J background. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J (Stock No. 000664) oocytes.

Allele

Symbol: Igs2^{tm1(CAG-AU040320/mCherry,-cas9*)Janc}
Name: targeted mutation 1, Jan Carette
MGI accession ID: MGI:7384598
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Synonyms: SELECTIV
Marker symbol: Igs2
Marker name: intergenic site 2
Marker synonyms: H11; Hipp11; Iis2
Chromosome: 11
Strain of origin: STOCK Igs2^tm3.1Luo/Luo
Molecular note: This allele was generated using PhiC31 integrase-mediated transgenesis. A plasmid containing from 5' to 3' a CAG promoter, a loxP-flanked stop sequence, mouse AU040320 (Aavr) fused to mCherry, an F2A self-cleaving peptide sequence, and a mammalian codon-optimized cas9 gene (derived from Streptococcus pyogenes CRISPR associated protein 9 [spCas9]) and PhiC31 integrase RNA was injected into the cytoplasm of C57BL/6 embryos carrying three tandem attP sites in the "safe harbor" Igs2 (H11) intergenic site, Igs2.
General note: This allele was produced in zygotes collected from mice carrying the Igs2^tm3.1Luo allele, which was generated in (129X1/SvJ x 129S1/Sv)F1-Kitl⁺-derived R1 ES cells on a mixed background with 129S1/SvJ, 129X1/SvJ, C57BL/6J, DBA/2J, and FVB/N.