Strain Detail Sheet

Strain Name:
STOCK Gt(ROSA)26Sortm1(CAG-cas9*,-BFP)Khk/Mmjax
Stock Number:
043926-JAX
Citation ID:
RRID:MMRRC_043926-JAX
Other Names:
Gt(ROSA)26Sor[tm1(CAG-dCas9-SunTag)Khk], Gt(ROSA)26Sor[tm1(CAG-dCas9-SunTag)Khk]

Strain Information

Gene Details

Gt(ROSA)26Sortm1(CAG-cas9*,-BFP)Khk
Name: gene trap ROSA 26, Philippe Soriano; targeted mutation 1, Klaus H Kaestner
Synonyms: Gt(ROSA)26Sor[tm1(CAG-dCas9-SunTag)Khk]
Type: Allele
Species: Multi-species
Chromosome: 6
Alteration at locus: Conditional
CAG
Name: CMV early enhancer/chicken beta actin promoter/rabbit beta globin splice acceptor
Type: DNA Segment
Species: Multi-species
Chromosome:
Alteration at locus: Conditional
Gt(ROSA)26Sor
Name: gene trap ROSA 26, Philippe Soriano
Synonyms: beta geo, R26, Gtrosa26, Thumpd3as1, Gtrgeo26, ROSA26
Type: Gene
Species: Mus musculus (mouse)
Chromosome: 6
Alteration at locus: Conditional
NCBI: 14910
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Genetic Alterations
Rosa26-LSL-dCas9 mice (Gt(ROSA)26Sortm1(CAG-dCas9-SunTag)Khk) are a Cre-inducible CRISPR/Cas9 activation (CRISPRa) tool for cell type specific gene activation in vivo.
Genotype Determination
ES Cell Line
v6.5

Strain Description

Phenotype
These Rosa26-LSL-dCas9 knock-in mice have a floxed-STOP cassette preventing expression of nuclease-deactivated dCas9 fused with the SunTag domain, and a BFP reporter under control of a CAG promoter. After exposure to Cre recombinase to remove the STOP cassette, dCas9/SunTag is expressed. The SunTag domain is made up of a peptide containing up to 24 copies of a short peptide epitope (GCN4) and a cognate single-chain antibody (scFv) that recognizes the epitope. When dCas9/SunTag is coexpressed with the antibody-transcriptional activator (TA) fusion, VP64, and a small guide RNA, the Tag allows for docking of multiple copies of the TA, VP64, to the target. Mice homozygous for the Rosa26-LSL-dCas9/SunTag knock-in allele are viable and fertile. When used in combination with single guide RNAs and a Cre source, these mice have applications in studies of in vivo, tissue specific modulation of gene expression and epigenetics and to perform scalable combinatorial genetic screens.
Of note, the blue fluorescent protein reporter (BFP) expression was not characterized by the donating laboratory.
Strain Development
The targeting vector contains (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-STOP-loxP followed by a catalytically inactive Cas9 protein (dCas9) fused to a repeating peptide array known as the SunTag domain, and a neomycin resistance gene under the control of the Pgk promoter inserted into intron 1 of the Rosa26 locus. The vector also contains a blue fluorescent protein reporter (BFP, however expression has not been characterized). The SunTag domain is made up of a peptide containing up to 24 copies of a short peptide epitope (GCN4) and a cognate single-chain antibody (scFv) that recognizes the epitope. When SunTag is fused to dCas9 and coexpressed with the antibody-transcriptional activator fusion, VP64, and a small guide RNA, the Tag allows for docking of multiple copies of VP64 to the target. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived v6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric male was crossed to a female B6.129S7-Fahtm1Mgo mouse. Offspring were crossed to mice on a B6SJLF1 background and the Fah allele was selected against. Mice were then crossed for one generation to C57BL/6J. Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony. This strain does not carry the Fahtm1Mgoallele.
Suggested Control Mice
Wild-type littermates

MMRRC Genetic QC

MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact csmmrrc@jax.org. Older strains may not have this information.

Research Applications

  • Cancer
  • Cardiovascular
  • Dermatology
  • Developmental Biology
  • Diabetes
  • Endocrine Deficiency
  • Hematology
  • Immunology and Inflammation
  • Internal_Organ
  • Metabolism
  • Models for Human Disease
  • Neurobiology
  • Obesity
  • Research Tools

Strain Origin

Donor
Kaestner [Ph.D.], Klaus
Primary Reference

Combinatorial genetics in liver repopulation and carcinogenesis with a novel in vivo CRISPR activation platform.Wangensteen KJ, Wang YJ, Dou Z, Wang AW, Mosleh-Shirazi E, Horlbeck MA, Gilbert LA, Weissman JS, Berger SL, Kaestner KH.Hepatology. 2017 Nov 1. doi: 10.1002/hep.29626. [Epub ahead of print]

Wangensteen KJ, Wang YJ, Dou Z, Wang AW, Mosleh-Shirazi E, Horlbeck MA,Gilbert LA, Weissman JS, Berger SL, Kaestner KH. Combinatorial genetics in liver repopulation and carcinogenesis with a novel in vivo CRISPR activation platform. Hepatology. 2017 Nov 1. doi: 10.1002/hep.29626. [Epub ahead of print] (Medline PMID: 29091290)

Colony and Husbandry Information

Special Considerations

None

Health Status Report

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-reports.

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email csmmrrc@jax.org.

Appearance

Coat Color
black

Breeding

MMRRC Breeding System
Backcross
Breeding Scheme(s)
no data
Generation
2
Overall Breeding Performance
Excellent

Reproductive Statistics

Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: Undetermined Undetermined
Bred to Homozygosity
Yes
Average litter size
5-9
Recommended wean age
3-4 weeks
Average Pups Weaned
5-9

Order Request Information

Availability Level

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

Conditions of Distribution

The donor or their institution limits the distribution to non-profit institutions only.

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

Fees

Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*		 Units Notes
043926-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
043926-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.

4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.