Strain Name:
B6J;129S4-Col1a1tm4Fcam/Mmjax
Stock Number:
067061-JAX
Citation ID:
RRID:MMRRC_067061-JAX
Other Names:
CARLIN

Strain Information

Col1a1tm4(CAG-EGFP)Fcam
Name: collagen, type I, alpha 1; targeted mutation 4, Fernando Camargo
Synonyms: Col1a1tm1(cCARLIN), CARLIN
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 11
Alteration at locus: Knock-In
Col1a1
Name: collagen, type I, alpha 1
Synonyms: Col1a-1, Mov-13, Cola-1, Cola1
Type: Gene
Species: Mouse
Chromosome: 11
Alteration at locus: Knock-In
NCBI: 12842
HGNC: HGNC:2197
Homologene: 73874
Genetic Alterations
The CARLIN (CRISPR array repair lineage tracing) allele was inserted into the 3' UTR of the Col1a1 locus. More specifically, CARLIN mice carry an expression cassette containing 10 guide RNAs (gRNA) with separate U6 promoters followed by a CAG-driven GFP reporter in which the sequence targeted by the gRNAs is inserted upstream of its polyadenylation site. When combined with doxycycline (dox), a dox-inducible allele expressing Cas9 (Col1a1tm1(tetO-cas9)Sho) and a tet reverse transactivator (Gt(ROSA)26Sortm1(rtTA*M2)Jae), cut and repair of the target sequence creates a unique and inheritable “barcode”. The target sequence was selected to allow RNA sequencing (RNAseq) of edited single cells thus allowing the determination of gene expression profiles and lineage history. Mice homozygous for the CARLIN allele are viable and fertile.

CARLIN is estimated to be able to generate 44,000 barcode arrays in an inducible fashion at any point in development or adulthood. Bowling et al. has created a database of barcode frequencies and analysis tools (PMID:32413320).

Genotype Determination
ES Cell Line
KH2 derived from (C57BL/6 x 129S4/SvJae)F1
Phenotype
None/Normal/Wild-type
Strain Development
The CARLIN components (10 constitutively expressed guide RNAs and CAG-GFP-CARLIN array sequence) were cloned into the pBS31 vector to target the Col1a1 locus in KH2 ESCs (derived from (C57BL/6 x 129S4/SvJae)F1). Hygromycin-selected clones were screened by PCR and injected into C57BL/6 embryos. Chimeras with >90% of chimerism (coat color) were used as founders, intercrossed to homozygosity, and occasionally backcrossed to C57BL/6.
Suggested Control Mice
Wild-type
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.
  • Cancer
  • Developmental Biology
  • Hematology
  • Research Tools
Donor
Fernando Camargo, Ph.D., Children's Hospital Boston.
Primary Reference

Bowling S, Sritharan D, Osorio FG, Nguyen M, Cheung P, Rodriguez-Fraticelli A,Patel S, Yuan WC, Fujiwara Y, Li BE, Orkin SH, Hormoz S, Camargo FD. AnEngineered CRISPR-Cas9 Mouse Line for Simultaneous Readout of Lineage Historiesand Gene Expression Profiles in Single Cells. Cell. 2020 May 13. pii:S0092-8674(20)30554-7. doi: 10.1016/j.cell.2020.04.048. [Epub ahead of print](Medline PMID: 32413320)

Colony and Husbandry Information

For more information about this colony's health status contact csmmrrc@jax.org
Coat Color
Black and agouti
Eye
Black
MMRRC Breeding System
Backcross and random intra-strain mating
Generation
N2+ (C57BL/6J), F?
Overall Breeding Performance
Excellent
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: 10 to 12 months Greater than 12 months
Bred to Homozygosity
Yes
Average litter size
7 to 9
Recommended wean age
4 Weeks
Average Pups Weaned
7 to 9

Order Request Information

Limited quantities of breeder mice (up to 2 males and 2 females or 4 mice) per investigator per month are available from a live colony, usually available to ship in under 12 weeks. Larger quantities may be available, please contact the distributing center directly at csmmrrc@jax.org for more details.

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

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.

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
067061-JAX-HOM-F
067061-JAX-HOM-M
Homozygous Female
Homozygous Male
$218.00 / Non-Profit Per Mouse The csmmrrc@jax.org may assess additional fees for any special requests (e.g., specific age or weight of mice, etc.).
067061-JAX-SPERM Cryo-preserved spermatozoa $437.00 / Non-Profit Aliquot Approximate quantity3
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.