H11Cas9 CRISPR/Cas9 knock-in mice have constitutive expression of CRISPR associated protein 9 (cas9) endonuclease directed by a CAG promoter. When used in with single guide RNAs, they allow editing of single or multiple mouse genes in vivo or ex vivo.
Monte M. Winslow, Stanford University School of Medicine
Genetic Background | Generation |
---|---|
?+pN6F4
|
Allele Type | Gene Symbol | Gene Name |
---|---|---|
Targeted (Endonuclease) | Igs2 | intergenic site 2 |
These H11Cas9 mice express the human codon-optimized S. pyogenes cas9 gene (hSpCas9) under control of a CAG promoter from the endogenous intergenic site 2, Igs2 locus (Hipp11 or H11; an intergenic region on chromosome 11 [cytoband A1 at ~3cM between the Eif4enif1 and Drg1 loci] reported to support high-level global gene expression in conjunction with the CAG (CAGGS) promoter. Whereas gene editing via viral delivery of cas9 is burdened by packaging size limits, these H11Cas9 mice only require delivery of a specific single guide RNA (sgRNA) for generating single or multiple simultaneous mutations. Mice homozygous for the H11Cas9 knock-in allele are expected to be viable and fertile.
In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for the strain above. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.
H11Cas9 knock-in mice are also available on a STOCK genetic background (Stock No. 027650).
A pattB-LSL-Cas9 plasmid and ΦC31 integrase mRNA were microinjected into the pronucleus and cytoplasm of H11attPx3/attPx3 zygotes on a 129 background. One founder mouse carried the correctly targeted transgenic insertion of pattB-LSL-Cas9 into the first and second attP sites of the H11attPx3 locus. The mouse was bred to 2 other mutant lines with B6;129 backgrounds. These H11LSL-Cas9 mice were bred to B6.C-Tg(CMV-cre)1Cgn/J (Stock No. 006054) mice to remove the floxed STOP cassette. The mice were then bred to remove the Cre recombinase transgene. The resulting H11Cas9 knock-in allele is therefore attL-CAGGS-loxP-3xFLAG-NLS-hSpCas9-NLS-polyA-attR-attP-FRT5. Male mice were sent to The Jackson Laboratory Repository in 2015 as Stock No. 027650. Upon arrival, some mice were then backcrossed to C57BL/6J inbred mice (Stock No. 000664) using a marker assisted protocol - the resulting C57BL/6J-congenic H11Cas9 strain is Stock No. 028239.
Expressed Gene | cas9, CRISPR associated protein 9, |
---|---|
Site of Expression |
Allele Name | targeted mutation 1.1, Monte Winslow |
---|---|
Allele Type | Targeted (Endonuclease) |
Allele Synonym(s) | H11Cas9 |
Gene Symbol and Name | Igs2, intergenic site 2 |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | cas9, CRISPR associated protein 9, |
Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl+ |
Chromosome | 11 |
Molecular Note | The targeting construct contains a full-length attB site, the CMV enhancer/chicken beta-actin core promoter (CAGGS), a loxP-STOP(3xSV40 polyA)- loxP cassette (LSL), a 3xFLAG sequence, a nuclear localization signal (NLS), a human codon-optimized S. pyogenes cas9 gene (hSpCas9), a second NLS and the SV40 polyA signal. The embryonic stem cells used were previously targeted with the H11 |
When maintaining a live colony, it is expected that these mice can be bred as homozygotes.
When using the H11Cas9 on B6J mouse strain in a publication, please cite the originating article(s) and include JAX stock #028239 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Igs2<tm1.1(CAG-cas9*)Mmw> |
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.