A copy number variation on human chromosome 16p11.2 is among the most common genetic variations found in autism spectrum disorders. 16p11flx mice are a Cre or FLP recombinase-inducible mouse model of 16p11.2 deletion that has several loxP and frt sites flanking the corresponding 440 kbp region on mouse chromosome 7F3. These mice may be useful in studying basal ganglia circuitry and the pathophysiology of autism.
Ricardo E Dolmetsch, Stanford University
Genetic Background | Generation |
---|---|
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), No functional change) | Igs13 | intergenic site 13 |
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Conditional ready (e.g. floxed), Reporter, No functional change) | Igs14 | intergenic site 14 |
A copy number variation (CNV) on human chromosome 16p11.2 is among the most common genetic variations found in autism spectrum disorders (ASD). Patients with this deletion display motor deficits, speech/language delay, and cognitive impairments, accompanied by ASD, attention deficit hyperactivity disorder (ADHD), seizures, and hearing disorders. Conversely, a duplication of 16p11.2 is associated with schizophrenia. The chromosome 16p11.2 CNV encompasses 26 genes that are highly conserved on mouse chromosome 7F3.
16p11flx is a Cre or FLP recombinase-inducible mouse model of 16p11.2 deletion that has several loxP and frt sites flanking the 440 kbp region on mouse chromosome 7F3. Homozygous mice (16p11flx) are viable and fertile with no reported gross physical or behavioral abnormalities.
Following Cre or FLP-mediated deletion of the entire region, a membrane-targeted fluorescent reporter gene (mCherry) is expressed. The donating investigator reports that for imaging large-scale anatomical structures, such as white matter tracts, mCherry fluorescence is very strong. For identification of single neuronal cell-bodies, however, immunohistochemical fluorescence might improve the outcome.
Mice with deletion of the genomic segment (16p11-) are available at The Jackson Laboratory as Stock No. 025100.
The 16p11flx model was created using two independent targeting events. Although the loxP/frt region near Coro1a is in relatively close proximity (440 kbp) to the loxP/frt region near Spn, the two loci have a small chance of segregating independently of one another. The donating investigator reports they never observed any crossing over between the two loci and they always segregated together.
Additionally, breeding heterozygous mice together at The Jackson Laboratory results in mice that are black, light chinchilla or albino. This suggests the 16p11flx mice we received retained mESC-derived allele(s) at the tyrosinase locus (e.g., Tyrc and/or Tyrc-ch) that, because it is only ~40 kbp proximal on chromosome 7, appears to be segregating along with the 16p11flx region. Depending upon future segregation patterns, available mice may be black, light chinchilla or albino.
16p11flx has several loxP and frt sites flanking the 440 kbp region on mouse chromosome 7F3; a region highly conserved with the human chromosome 16p11.2 region whose copy number variations are associated with autism spectrum disorders and schizophrenia.
Drs. Ricardo E. Dolmetsch and Thomas Portmann (while at Stanford University) created the two targeting vectors used to generate these 16p11flx mice. The first vector was designed to insert a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), loxP-frt-neomycin resistance cassette-loxP-frt-loxP sequence and polyA signal at the centromeric end of the coronin actin binding protein 1A locus (Coro1a). The insertion site was 1165 bp centromeric (transcriptionally downstream) of the last Coro1a exon to avoid disrupting endogenous expression prior to Cre recombination. The second targeting vector was designed to insert a translational stop codon (for all frames), loxP-frt-puromycin resistance cassette-loxP-frt sequence, rabbit b-globin intron, coding sequence mCherry fluorescent protein (with palmitoylation signal for membrane targeting) and polyA signal at the telomeric end of the sialophorin locus (Spn). The insertion site was 3336 bp telomeric (transcriptionally upstream) of the Spn gene to avoid disrupting endogenous expression prior to Cre recombination. Sequential targeting into 129 mouse embryonic stem cells (mESC [see note below]) resulted in mESC clones with cis arrangement (targeting into the same chromosome 7 homolog) or trans arrangement (each vector targeted into different chromosome 7 homologs). The modified mESCs were injected into blastocysts, implanted into pseudopregnant female mice, and chimeric males were subsequently bred to C57BL/6NCrl females and/or HPRT-Cre females (Hprttm1(cre)Mnn on a C57BL/6;CD1 mixed genetic background; see Stock No. 004302). Germline transmission was successful only for the cis arrangement, and animals with no recombination resulted in heterozygous floxed mice (16p11flx/+). The 16p11flx colony was bred to C57BL/6NCrl for at least five generations prior to sending black males to The Jackson Laboratory in 2014. Upon arrival, sperm was cryopreserved. To establish our living colony, an aliquot of frozen sperm was used to fertilize C57BL/6NJ oocytes (from Stock No. 005304).
Additionally, breeding heterozygous mice together at The Jackson Laboratory results in mice that are black, light chinchilla or albino. While the Portmann et al. 2014 Cell Rep 7(4):1077-92 publication describes the mESC used were 129/OLA, the donating laboratory indicates it may have been a different 129 origin. Taken together, these data suggest the 16p11flx mice we received retained mESC-derived allele(s) at the tyrosinase locus (e.g., Tyrc and/or Tyrc-ch) that, because it is only ~40 kbp proximal on chromosome 7, appears to be segregating along with the 16p11flx region. Depending upon future segregation patterns, available mice may be black, light chinchilla or albino.
In 2015, a 32 SNP panel analysis (27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains), was performed on the first generation rederived live colony at The Jackson Laboratory Repository. This revealed one animal with 2 of 27 markers, one each on chromosomes 2 and 12, that were not fixed for C57BL/6 allele-type (e.g., still segregating for 129 allele-type markers). All 5 markers that determine C57BL/6 substrains were found to be C57BL/6N allele-type.
Expressed Gene | RFP, Red Fluorescent Protein, coral |
---|---|
Site of Expression | Following Cre or FLP-mediated deletion of the 440 kbp region on mouse chromosome 7F3region, a membrane-targeted fluorescent reporter gene (mCherry) is expressed in Cre or FLP expressing tissues. |
Allele Name | targeted mutation 1, Ricardo Dolmetsch |
---|---|
Allele Type | Targeted (Conditional ready (e.g. floxed), No functional change) |
Allele Synonym(s) | |
Gene Symbol and Name | Igs13, intergenic site 13 |
Gene Synonym(s) | |
Site of Expression | Following Cre or FLP-mediated deletion of the 440 kbp region on mouse chromosome 7F3region, a membrane-targeted fluorescent reporter gene (mCherry) is expressed in Cre or FLP expressing tissues. |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 7 |
Molecular Note | The targeting vector is designed to insert a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), loxP-frt-neomycin resistance cassette-loxP-frt-loxP sequence and polyA signal at the centromeric end of the coronin actin binding protein 1A locus (Coro1a) gene. The insertion site is 1165 bp centromeric (transcriptionally downstream) of the last exon. |
Allele Name | targeted mutation 1, Ricardo Dolmetsch |
---|---|
Allele Type | Targeted (Conditional ready (e.g. floxed), Reporter, No functional change) |
Allele Synonym(s) | |
Gene Symbol and Name | Igs14, intergenic site 14 |
Gene Synonym(s) | |
Expressed Gene | RFP, Red Fluorescent Protein, coral |
Site of Expression | Following Cre or FLP-mediated deletion of the 440 kbp region on mouse chromosome 7F3region, a membrane-targeted fluorescent reporter gene (mCherry) is expressed in Cre or FLP expressing tissues. |
Strain of Origin | 129P2/OlaHsd |
Chromosome | 7 |
Molecular Note | The targeting vector is designed to insert a translational stop codon (for all frames), loxP-frt-puromycin resistance cassette-loxP-frt sequence, rabbit b-globin intron, coding sequence mCherry fluorescent protein (with palmitoylation signal for membrane targeting) and polyA signal at the telomeric end of the sialophorin locus (Spn) gene. The insertion site is 3336 bp telomeric (transcriptionally upstream). |
When maintaining a live colony, mice homozygous for each targeted allele may be bred together.
Additionally, breeding heterozygous mice together at The Jackson Laboratory results in mice that are black, light chinchilla or albino. While the Portmann et al. 2014 Cell Rep 7(4):1077-92 publication describes the mESC used were 129/OLA, the donating laboratory indicates it may have been a different 129 origin. Taken together, these data suggest the 16p11flx mice we received retained mESC-derived allele(s) at the tyrosinase locus (e.g., Tyrc and/or Tyrc-ch) that, because it is only ~40 kbp proximal on chromosome 7, appears to be segregating along with the 16p11flx region. Depending upon future segregation patterns, available mice may be black, light chinchilla or albino.
When using the 16p11.2flx mouse strain in a publication, please cite the originating article(s) and include JAX stock #025330 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous or Wildtype for Igs13<tm1Dolm> and for Igs14<tm1Dolm> |
Frozen Mouse Embryo | B6N.129P2(Cg)-Igs13<tm1Dolm> Igs14<tm1Dolm>/J | $2595.00 |
Frozen Mouse Embryo | B6N.129P2(Cg)-Igs13<tm1Dolm> Igs14<tm1Dolm>/J | $2595.00 |
Frozen Mouse Embryo | B6N.129P2(Cg)-Igs13<tm1Dolm> Igs14<tm1Dolm>/J | $3373.50 |
Frozen Mouse Embryo | B6N.129P2(Cg)-Igs13<tm1Dolm> Igs14<tm1Dolm>/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.