Strain Name:
C57BL/6J-Gt(ROSA)26Sorem2Mvw/MvwJ
Stock Number:
069681-JAX
Citation ID:
RRID:MMRRC_069681-JAX
Other Names:
C57BL/6J-Gt(ROSA)26Sorem2Mvw/MvwJ

Strain Information

Gt(ROSA)26Sorem2Mvw
Name: gene trap ROSA 26, Philippe Soriano; endonuclease-mediated mutation 2, Michael Wiles
Synonyms: B6.RosaBxb-GT
Type: Allele
Species: Mus musculus
Chromosome: 6
Alteration at locus: CRISPR
Gt(ROSA)26Sor
Name: gene trap ROSA 26, Philippe Soriano
Synonyms: ROSA26, beta geo, Gtrgeo26, Gtrosa26, R26, Thumpd3as1
Type: Gene
Species: Mus musculus (mouse)
Chromosome: 6
Alteration at locus: CRISPR
NCBI: 14910
Genetic Alterations
RosaBxb-GT mice are a resource for the rapid creation of vector-free targeted alleles in the Gt(ROSA)26Sor locus on the C57BL/6J background. The single Bxb1 integrase attP-site "safe harbor landing pad" in the first intron of Gt(ROSA)26Sor enables efficient and precise integration of a donor DNA sequence directly in zygotes by recombinase-mediated knock-in (RMKI) using vector-free minicircle donor DNA carrying the transgene of interest and the cognate attB-GT attachment site.
Genotype Determination
ES Cell Line
Not applicable
Phenotype
This “safe harbor landing pad” allele on the C57BL/6J background provides a tool for precise, site-specific insertion of single-copy sequence into the Gt(ROSA)26Sor intron 1 in a manner that is in the desired orientation and free from vector sequence. In practice, a plasmid containing a single cognate 38 bp attB-GT site and the donor sequence is first converted into a vector-less DNA minicircle donor. The DNA minicircle is delivered along with the Bxb1 integrase (mRNA or protein) into the pronucleus of fertilized oocytes containing the 48 bp attP-GT landing pad attachment site. The result is a unidirectional, single-copy integration of the sequence into the Gt(ROSA)26Sor locus. The inserted sequence will be flanked by the recombined attachments sites, now called Bxb1 attL and Bxb1 attR, but will be: (1) free from residual vector sequence from the plasmid backbone, (2) present as a single-copy (no concatemers), and (3) inserted precisely into a known and verifiable locus (Gt(ROSA)26Sor). [Low et al., 2022] The donating investigator indicates that 5% of integrations result in aberrant integration events. In addition, RMKI strategy is successful with inserts up to approximately 15 kb. Please note that this strain carries a +6 bp/-3 bp indel preceding the 5’ end of the attP-GT site.

Both heterozygous and homozygous mice are viable and fertile with normal breeding, and no reported gross phenotypic or behavioral abnormalities.
MeSH Terms
  • Animals
  • Bacteriophages/genetics
  • COVID-19
  • DNA
  • Gene Transfer Techniques
  • Genetic Background
  • Integrases/genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred DBA
  • Mice, Inbred NOD
  • RNA, Viral
  • SARS-CoV-2
Strain Development
The RosaBxb-GT allele was generated in the laboratory of Dr. Michael V. Wiles (The Jackson Laboratory) using CRISPR/Cas9 endonuclease-mediated genome editing/oligo-mediated homology directed repair. An 18-mer TRU-guide RNA was selected to target near the XbaI site of intron 1 in the Gt(ROSA)26Sor locus on Chromosome 6. A 200 nt ssDNA donor (with 76 nt homology arms flanking the 48 bp Bxb1 attP-GT attachment site), the TRU-guide and Cas9 mRNA were introduced into C57BL/6J zygotes by microinjection. Progeny were screened by PCR amplification and DNA sequencing to identify correctly targeted pups. Sequencing identified a +6 bp/-3 bp indel preceding the 5' end of the attP-GT site. Founder mice were then backcrossed at least once to C57BL/6J mice for germline transmission and allele verification before intercrossing to generate homozygotes.
Suggested Control Mice
C57BL/6J
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 Tools
Donor
Aamir Zuberi, Ph.D., The Jackson Laboratory.
Michael Wiles, The Jackson Laboratory.
Primary Reference

Low BE, Hosur V, Lesbirel S, Wiles MV. Efficient targeted transgenesis of large donor DNA into multiple mouse genetic backgrounds using bacteriophage Bxb1 integrase. Sci Rep. 2022 Mar 31;12(1):5424. doi: 10.1038/s41598-022-09445-w. (Medline PMID: 35361849)

Colony and Husbandry Information

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.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
N/A
Overall Breeding Performance
Undetermined
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
Average litter size
Undetermined
Recommended wean age
4 Weeks
Average Pups Weaned
Undetermined

Order Request Information

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.

A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Peter Wells.

A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Peter Wells

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.

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
069681-JAX-SPERM Cryo-preserved spermatozoa $437.00 / $437.00
Non-Profit / For-Profit
Aliquot Approximate quantity3
069681-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / $2,022.00
Non-Profit / For-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.