Overview

Also Known As:H11, H11^LSL-Cas9

H11^LSL-Cas9 CRISPR/Cas9 knock-in mice have Cre recombinase-dependent expression of CRISPR associated protein 9 (cas9) endonuclease directed by a CAG promoter. When used in combination with single guide RNAs and a Cre recombinase source, they allow editing of single or multiple mouse genes in vivo or ex vivo.

Donating Investigator

Monte M. Winslow, Stanford University School of Medicine

Genetic overview

Genetic Background	Generation
	N5F12 (2020-12-07 00:00:00)

Igs2^{tm1(CAG-cas9)Mmw}*

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), No functional change, Endonuclease)	Igs2	intergenic site 2

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Research Applications

Research Tools

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Base Price

Starting at:

$305.90 Domestic price for female 4-week

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Details

Detailed Description

These H11^LSL-Cas9 knock-in mice have a loxP-flanked STOP cassette preventing expression of the downstream sequence (cas9), inserted in the 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 and a higher rate of mitotic/interchromosomal recombination compared to Gt(ROSA)26Sor). Although under control of a CAG promoter, widespread expression of cas9 is prevented by the STOP cassette. After exposure to Cre recombinase, expression of cas9 is observed. Whereas gene editing via viral delivery of cas9 is burdened by packaging size limits, these H11^LSL-Cas9 mice only require the user to select a Cre recombinase driven by the promoter of their choosing and a specific single guide RNA (sgRNA) for generating single or multiple simultaneous mutations. Mice homozygous for the H11^LSL-Cas9 knock-in allele are viable and fertile.

H11^LSL-Cas9 knock-in mice are also available on a B6;129 genetic background (Stock No. 026816).

Development

The H11^LSL-Cas9 knock-in allele has attL-pCA-LSL-3xFLAG-NLS-hSpCas9-NLS-polyA-attR-attP-FRT5 inserted into the 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 and a higher rate of mitotic/interchromosomal recombination compared to Gt(ROSA)26Sor). Specific details below.
The pattB-LSL-Cas9 targeting vector was created with 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. (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells were previously targeted with the H11^attPx3 allele (Igs2^tm3.1Luo or H11P3; a unique sequence from the promoter of the yeast his3 gene followed by three shortened tandem attP sites and one FRT5 site). H11^attPx3 mice were backcrossed onto C57BL/6, and then made homozygous (H11^{attPx3/attPx3}). For site-specific ΦC31 integrase-mediated transgenesis, the pattB-LSL-Cas9 construct and ΦC31 integrase mRNA were microinjected into the pronucleus and cytoplasm of H11^{attPx3/attPx3} zygotes. One founder mouse carried the correctly targeted insertion of pattB-LSL-Cas9 into the first and second attP sites of the H11^attPx3 locus. The resulting H11^LSL-Cas9 knock-in allele is therefore attL-CAGGS-LSL-3xFLAG-NLS-hSpCas9-NLS-polyA-attR-attP-FRT5. The H11^LSL-Cas9LSL-Cas9 colony was bred to 2 other mutant lines with B6;129 backgrounds prior to sending agouti males to The Jackson Laboratory Repository in 2015 as Stock No. 026816. 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 H11^LSL-Cas9 strain is Stock No. 027632.

Expression Data

Expressed Gene	cas9, CRISPR associated protein 9,
Site of Expression

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Chiou SH; Winters IP; Wang J; Naranjo S; Dudgeon C; Tamburini FB; Brady JJ; Yang D; Gruner BM; Chuang CH; Caswell DR; Zeng H; Chu P; Kim GE; Carpizo DR; Kim SK; Winslow MM. 2015. Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing. Genes Dev 29(14):1576-85PubMed: 26178787 MGI: J:222466

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Genetics

Igs2^{tm1(CAG-cas9)Mmw}*

Allele Symbol: Igs2^{tm1(CAG-cas9)Mmw}*

Allele Name	targeted mutation 1, Monte Winslow
Allele Type	Targeted (Conditional ready (e.g. floxed), No functional change, Endonuclease)
Allele Synonym(s)	H11^LSL-Cas9
Gene Symbol and Name	Igs2, intergenic site 2
Gene Synonym(s)
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 allele ( Igs2^tm3.1Luo) or H11P3; a unique sequence from the promoter of the yeast his3 gene followed by three shortened tandem attP sites and one FRT5 site. One founder mouse carried the correctly targeted insertion of pattB-LSL-Cas9 into the first and second attP sites of the H11 locus. The resulting H11 knock-in allele is therefore attL-CAGGS-LSL-3xFLAG-NLS-hSpCas9-NLS-polyA-attR-attP-FRT5.

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Cre-lox System
  - loxP-flanked Sequences
- CRISPR/Cas9 system

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Igs2^{tm1(CAG-cas9)Mmw}/Igs2^{tm1(CAG-cas9)Mmw}
involves: 129S1/Sv * 129X1/SvJ * C57BL/6

normal phenotype

no abnormal phenotype detected
- mice are viable and fertile

References

Chiou SH; Winters IP; Wang J; Naranjo S; Dudgeon C; Tamburini FB; Brady JJ; Yang D; Gruner BM; Chuang CH; Caswell DR; Zeng H; Chu P; Kim GE; Carpizo DR; Kim SK; Winslow MM. 2015. Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing. Genes Dev 29(14):1576-85PubMed: 26178787 MGI: J:222466

Additional - Igs2^{tm1(CAG-cas9)Mmw}* related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Igs2
- Probe:Igs2 Probe
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, these mice can be bred as homozygotes.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the H11^LSL-Cas9 mouse strain in a publication, please cite the originating article(s) and include JAX stock #027632 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX12 (Maximum)

Pricing & Availability

Available

Domestic
International

Domestic Pricing for

Commercial & For-Profit

Not-For-Profit & Academic

International Pricing for

Commercial & For-Profit

Not-For-Profit & Academic

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous or wildtype for Igs2<tm1(CAG-cas9*)Mmw>

Related Products and Services

Frozen Mouse Embryo	B6.129-Igs2<tm1(CAG-cas9*)Mmw>/J	$2595.00
Frozen Mouse Embryo	B6.129-Igs2<tm1(CAG-cas9*)Mmw>/J	$2595.00
Frozen Mouse Embryo	B6.129-Igs2<tm1(CAG-cas9*)Mmw>/J	$3373.50
Frozen Mouse Embryo	B6.129-Igs2<tm1(CAG-cas9*)Mmw>/J	$3373.50

Payment Terms and Conditions

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's Genotype Promise

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.

Terms Of Use

General Terms and Conditions

Questions about Terms of Use

Additional Use Restrictions Apply

Use of MICE by companies or for-profit entities requires a license prior to shipping.

Licensing Information

Phone: 207-288-6470

Email: TechTran@jax.org

Also Known As:H11, H11LSL-Cas9

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Igs2tm1(CAG-cas9*)Mmw

Allele Symbol: Igs2tm1(CAG-cas9*)Mmw

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Igs2tm1(CAG-cas9*)Mmw/Igs2tm1(CAG-cas9*)Mmwinvolves: 129S1/Sv * 129X1/SvJ * C57BL/6

normal phenotype

References

Additional - Igs2tm1(CAG-cas9*)Mmw related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

Also Known As:H11, H11^LSL-Cas9

Igs2^{tm1(CAG-cas9)Mmw}*

Allele Symbol: Igs2^{tm1(CAG-cas9)Mmw}*

Genotype: Igs2^{tm1(CAG-cas9)Mmw}/Igs2^{tm1(CAG-cas9)Mmw}
involves: 129S1/Sv * 129X1/SvJ * C57BL/6

Additional - Igs2^{tm1(CAG-cas9)Mmw}* related