Strain Name:
B6J;B6N-Espnlem1Pgg/Mmucd
Stock Number:
050613-UCD
Citation ID:
RRID:MMRRC_050613-UCD
Other Names:
Espnl, Espn-like, EspnlΔ

Strain Information

Espnlem1Pgg
Name: espin-like; endonuclease-mediated mutation 1, Peter G Gillespie
Synonyms: Espnldelta
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 1
Alteration at locus: CRISPR
Espnl
Name: espin-like
Synonyms: LOC227357
Type: Gene
Species: Mouse
Chromosome: 1
Alteration at locus: CRISPR
NCBI: 227357
Homologene: 77795
Genetic Alterations
CRISPR-mediated targeting of the Espnl gene (exons 1 and 8). More specifically:

HGVS nomenclature:
  • Genbank RefSeq - mRNA: NM_001033292.3
  • Genbank RefSeq, protein: NP_001028464.1
  • Variant #1, nucleic acid level: c.136_1300del
  • Variant #1, amino acid level, predicted: p.Ala46Thrfs*14
    Check this variant: LUMC Mutalyzer
Phenotype
High-frequency hearing loss, abnormal stereocilia in outer cochlear hair bundles
MeSH Terms
  • Animals
  • COS Cells
  • Cercopithecus aethiops
  • Ear, Inner/metabolism
  • Female
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Microfilament Proteins/genetics
  • Microfilament Proteins/metabolism
  • Myosin Heavy Chains/genetics
  • Myosin Heavy Chains/metabolism
  • Myosin Type III/genetics
  • Myosin Type III/metabolism
  • Rats
  • Stereocilia/physiology
  • Tissue Culture Techniques
Strain Development
For the Espnl knockout with CRISPR, gRNAs targeted to Espnl exons 1 and 8 were selected using the CRISPR web-based tool (http://crispr.mit.edu/). The gRNA sequences (exon 1, 5′-GTGCATCATGCCACCCGGGC-3′; exon 8, 5′-CCGGCCACGCTCGTCCTGTG-3′) were individually cloned into the DR274 plasmid (Addgene #42250) and transcribed using the MegaScript kit (LifeTechnologies). gRNAs were purified using the NucleoSpin miRNA (Macherey-Nagel) and quantified. C57BL/6N zygotes were injected with a cocktail containing 30 ng μl−1 of each gRNA and 110 ng μl−1 of Cas9 mRNA (Trilink) and implanted into pseudopregnant females. All founders were backcrossed to N3 (C57BL/6N) and intercrossed to homozygosity (F9); two backcrosses to C57BL/6J prior to submission to the MMRRC. Screening for mutations in Espnl exon 1, exon 8, and any intervening deletions between these two exons, as well as all genes with predicted off-target sites (exon 1: AW551984, C1ql1, Cyp26c1, Olfr523, Pdc4c, Pomt2, and Slc12a7; exon 8: Acss2, Elf4, Krt79, Lrrc30, Ncoa4 and Phf2) in the founder mice demonstrated that all off-target sites were wild-type.

Note: The publication refers to c.136_1300del (corrected from c.135_1301del1165), predicting protein alteration p.Ala46Thrfs*14, as EspnlΔ.
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 mmrrc@ucdavis.edu. Older strains may not have this information.
  • Neurobiology
  • Sensorineural
Donor
Peter Barr-Gillespie, Ph.D., Oregon Health & Science University.
Primary Reference

Ebrahim S, Avenarius MR, Grati M, Krey JF, Windsor AM, Sousa AD, BallesterosA, Cui R, Millis BA, Salles FT, Baird MA, Davidson MW, Jones SM, Choi D, Dong L, Raval MH, Yengo CM, Barr-Gillespie PG, Kachar B. Stereocilia-staircase spacing isinfluenced by myosin III motors and their cargos espin-1 and espin-like. NatCommun. 2016 Mar 1;7:10833. doi: 10.1038/ncomms10833. Erratum in: Nat Commun.2017 Aug 14;8:16133. (Medline PMID: 26926603)

Colony and Husbandry Information

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.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
N2 (C57BL/6J), F9
Overall Breeding Performance
Good
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: 6 to 8 months Undetermined
Bred to Homozygosity
Yes
Average litter size
4-6
Recommended wean age
3 Weeks
Average Pups Weaned
4-6

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 mmrrc@ucdavis.edu for more information.

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
050613-UCD-SPERM Cryo-preserved spermatozoa $546.25 / $869.88
Non-Profit / For-Profit
Aliquot Approximate quantity3
050613-UCD-RESUS Litter recovered from cryo-archive $4,044.00 / $7,650.23
Non-Profit / For-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.