B6.Cg-Commd10^{Tg(Vav1-icre)A2Kio} Tg(CAG-glnK1*/mVenus)1Bous/GrandJ

Stock number: 041913
Product name: Vav-iCre-SPICY
Research areas: Cancer Research, Immunology, Inflammation and Autoimmunity Research, Metabolism Research, Research Tools

Description

Vav-iCre-SPICY mice express both a hematopoietic cell specific Cre (Vav1-icre) and a transgene harboring a loxP-flanked PercevalHR, an ATP:ADP fluorescent biosensor. These mice may be a useful tool for tracking individual hematopoietic cells and their respective metabolic profile using the SPICE-Met (Single-cell Profiling and Imaging of Cell Energy Metabolism) method.

Detailed description

PercevalHR (Perceval High Resolution) is an artificially designed fluorescent protein, which changes its excitation spectrum based on the ADP/ATP ratio of the environment. Vav-iCre-SPICY mice express both a hematopoietic cell specific Cre (Vav1-icre) and a transgene harboring a Cre-inducible PercevalHR.

Mice that are hemizygous for the Vav1-icre allele and hemizygous for the PervecalHR transgene are viable and fertile with no overt phenotype. The donating laboratory has not attempted to generate mice homozygous for the PervecalHR transgene. Homozygous Commd10^{Tg(Vav1-icre)A2Kio} mice are not viable (Stock No. 008610). mVenus expression was detected by flow cytometry in T cells, B cells, NK cells, monocytes, and neutrophils. These mice may be a useful tool for tracking individual hematopoietic cells and their respective metabolic profile using the SPICE-Met (Single-cell Profiling and Imaging of Cell Energy Metabolism) method.

Of note, Joseph et al. 2013 Cell Stem Cell 13:520 reports that Vav1-icre mice express iCre recombinase in hematopoietic cells and endothelial cells, as well as in the testes. For Cre-lox experiments and to avoid/minimize germline deletion of the floxed allele, researchers may consider breeding Vav-icre females with floxed males. However, female germline expression may also be observed (MGI:2449949).

Development

The Vav-iCre-SPICY double mutant mouse model was generated by breeding B6.Cg-Commd10Tg^{(Vav1-icre)A2Kio}/J (Stock No. 008610) to a conditional transgenic SPICY (PervecalHR^fl) mouse line. Additional details described below.

The conditional SPICY transgene expresses PercevalHR (GW1-Perceval from Gary Yelen, Addgene #49082), a synthetic fluorescent biosensor that was generated by mutating the original Perceval sensor at and around the GlnK nucleotide binding site. The final transgene construct includes a loxP-flanked STOP (PGK-Neo) cassette, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), the PercevalHR coding sequence, harboring a mutant GlnK site and circular permuted monomeric yellow fluorescent protein (mVenus) and a polyA tail. The purified linearized DNA was microinjected into the pronuclei of fertilized C57BL/6JOlaHsd mouse oocytes. Founder line 1 was determined to harbor the loxP-flanked PercevalHR cassette and further expanded. The transgenic copy number and integration site have not been evaluated. The conditional SPICY (Pervecal^fl) mice were then bred to the Vav1-icre mouse line (Stock No. 008610).

The Vav-iCre transgene was designed with the mouse vav 1 oncogene (Vav1) promoter region (containing the 2 proximal upstream DNase-I hypersensitive (HS) sites (HS21)), an optimized variant of Cre recombinase (iCre; improved with mammalian codon usage, removed putative cryptic splice sites, altered stop codon, and reduced CpG content to limit the chances of epigenetic silencing in mammals) and an SV40 polyA signal replacing Vav1 exon 1, and Vav1 intron 1 (containing both proximal HS sites (HS45)). The transgene was microinjected into the pronuclei of fertilized oocytes from (CBA/Ca x C57BL/10)F1 mice. Founder mice (founder line ".A2") were established and used to generate mutant mice. The resulting Vav-iCre transgenic mice were subsequently bred to wildtype C57BL/6 mice for more than 16 generations prior to arrival at The Jackson Laboratory Repository. In 2018, it was discovered that the transgene integrated into an intron of Commd10 (COMM domain containing 10) on chromosome 18 - creating a Commd10 null allele. The C57BL/6-congenic Vav-iCre colony is Stock No. 008610.

Dr. Philippe Bousso acquired some mice from Stock No. 008610 and bred them to the conditional SPICY (Pervecal^fl) mice to generate this Vav-iCre-SPICY double mutant mouse model. This mouse line was then transferred to Dr. Capucine Grandjean and maintained by intercrossing Vav1-icre hemizygous mice with conditional SPICY (Pervecal^fl) hemizygous mice to generate the Vav-iCre-SPICY double mutant mouse line. Upon arrival at The Jackson Laboratory, 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: Commd10^{Tg(Vav1-icre)A2Kio}
Name: transgene insertion A2, Dimitris Kioussis
MGI accession ID: MGI:2449949
Generation method: Transgenic
Attributes: Recombinase-expressing; Null/Knockout
Synonyms: Tg(Vav1-cre)1Kio; Tg(Vav1-cre)A2Kio; Tg(Vav1-icre)A2Kio; Vav1 (HS21/5)-iCre; Vav1-Cre; VavCre; Vav-cre; VavCre^deBoer; Vav-iCre
Marker symbol: Commd10
Marker name: COMM domain containing 10
Marker synonyms: 2310003A05Rik; Arfip2-ps1; DRWMS2; PTD002
Chromosome: 18
Strain of origin: (CBA/Ca x C57BL/10)F2
Expressed genes: cre,cre recombinase,bacteriophage P1
Site of expression: Cre recombinase is expressed in lymph nodes, thymus, spleen, Peyer's patches, intestinal cryptopatches, ovaries and testes.
Molecular note: This transgene expresses Cre recombinase under the control of a vav promoter. This promoter was shown to be active in lymph nodes, thymus, spleen, Peyer's patches, intestinal cryptopatches, ovaries and testes. Line A2 inserted into the gene at 47022629-47022630 (Build GRCm38/mm10). The insertion results in a functional knock-out in homozygous mice.

Allele

Symbol: Tg(CAG-glnK1*/mVenus)1Bous
Name: transgene insertion 1, Philippe Bousso
MGI accession ID: MGI:8403113
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Synonyms: PervecalHR^fl
Marker symbol: Tg(CAG-glnK1*/mVenus)1Bous
Marker name: transgene insertion 1, Philippe Bousso
Marker synonyms: Tg(CAG-Glnk1*,-mVenus)1Bous
Chromosome: UN
Strain of origin: C57BL/6JOlaHsd
Molecular note: The transgene expresses PercevalHR, a synthetic fluorescent biosensor that was generated by mutating the original Perceval sensor at and around the GlnK nucleotide binding site. The final transgene construct includes a loxP-flanked STOP (PGK-Neo) cassette, a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), the PercevalHR coding sequence harboring a mutant bacterial glnK1 sequence, circular permuted monomeric yellow fluorescent protein (mVenus), and a polyA tail.