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Strain Detail Sheet

Strain Name:
STOCK MARC1-PB7/Mmucd
Stock Number:
065424-UCD
Citation ID:
RRID:MMRRC_065424-UCD
Other Names:
MARC1

Strain Information

Gene Details

Genetic Alterations

Insertion of 60 homing guide RNA (hgRNA) loci in locations scattered throughout the genome. Each mouse will contain a subset of the inserted homing guide RNA's and the MMRRC will provide a report with each mouse containing the details on the inserted homing guides. Lineage barcoding experiments can be carried out by crossing these mice to a line that expressesCas9 constituitively or conditionally. In progeny of such a cross, hgRNAs begin to mutate themselves in cells that express Cas9 and accumulate heritable mutations.The cells or cell populations under investigation can then be isolated, their hgRNAs amplified and sequenced to obtain their lineage barcodes. Comparison between barcodes of different samples within the same mouse can shed light on the lineage relationship between those samples. Please consult the associated publication (PMID:30093604) for details.

Genotype Determination
An additional resource for analysis is: https://github.com/Kalhor-Lab/MARC1-Pipeline.
ES Cell Line
v6.5 derived from (C57BL/6 x 129S4/SvJae)F1

Strain Description

Phenotype
This mouse line is called "Mouse For Actively Recording Cells" or MARC1. It is used for barcoding and lineage tracing applications in the mouse, as described in the referenced publications. The line does not have a disease phenotype. However, when a MARC1-PB7 mouse is crossed to mice expressing cas9 universally or in a lineage-specific manner, combinatorial and cumulative barcoding starts in the progeny during their development, leading to developmentally barcoded animals that can be used for a multitude of applications, including lineage tracing. The MARC1-PB7 line was founded by a chimeric male mouse that carried 60 homing guide RNA (hgRNA) loci scattered throughout its genome in a heterozygous state. The founder (#7) was crossed to wild-type mice two years prior to archiving and has been inbred since with the goal of obtaining a line that is homozygous for all 60 loci.
MeSH Terms
  • Alleles
  • Animals
  • CRISPR-Cas Systems
  • Cell Lineage
  • Embryonic Development/genetics
  • Embryonic Stem Cells
  • Gene Expression Profiling/methods
  • Mice
  • Mutation
  • RNA, Guide/genetics
Strain Development
A barcoded library of homing guide RNAs (hgRNAs) with randomized spacer sequences was transposed in v6.5 mESCs using the Piggbac system. Transposed mESCs were selected using puromycin, injected into blastocysts, implanted in surrogate females at the Harvard Genome Modification Core Facility. Among the resulting chimeric mice, males with the highest number of hgRNA integrations were selected as founders of the MARC1 line. One of these founders (Chimera #7) carried 60 heterozygote germline-transmissible integrations of hgRNAs. The selected founders were crossed with B6 or CD1 females to create the F1 generation. In the F1 generation, PCR amplification of the hgRNA amplicon followed by high throughput sequencing was used to determine the number of hgRNAs inherited to each member. Based on these numbers, pairs of F1 were selected with the objective of maximizing the number of hgRNAs that would be inherited to F2, and crossed to create F2. The same procedure was repeated for F2 and so on to create the following generations. On average, 50 mice were generated per generation, genotyped (by PCR amplification followed by Illumina sequencing), and used to create the next generation.

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 mmrrc@ucdavis.edu. Older strains may not have this information.

Research Applications

  • Cancer
  • Cardiovascular
  • Developmental Biology
  • Diabetes
  • Hematology
  • Immunology and Inflammation
  • Internal/Organ
  • Models for Human Disease
  • Neurobiology
  • Obesity
  • Research Tools

Strain Origin

Donor
George M. Church, Ph.D., Harvard Medical School.
Reza Kalhor, Ph.D., Harvard Medical School.
Primary Reference

Kalhor R, Kalhor K, Mejia L, Leeper K, Graveline A, Mali P, Church GM.Developmental barcoding of whole mouse via homing CRISPR. Science. 2018 Aug31;361(6405). pii: eaat9804. doi: 10.1126/science.aat9804. Epub 2018 Aug 9.(Medline PMID: 30093604)

Colony and Husbandry Information

Special Considerations

Strain recovery fees do not include genetic analysis.

Health Status Report

Colony Surveillance Program and Current Health 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 mmrrc@ucdavis.edu.

Microbiome Analysis

Appearance

Coat Color
Mixed
Eye
Other

Breeding

MMRRC Breeding System
Random intra-strain mating
Generation
F10
Overall Breeding Performance
Good

Reproductive Statistics

NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Undetermined Undetermined
Hetero/Hemizygotes are fertile: Undetermined Undetermined
Age Reproductive Decline: Variable Variable
Bred to Homozygosity
No
Average litter size
4-6
Recommended wean age
3 Weeks
Average Pups Weaned
4-6

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 mmrrc@ucdavis.edu 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
065424-UCD-RESUS Litter recovered from cryo-archive $4,044.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to mmrrc@ucdavis.edu 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.