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Actg1c-b mutation knock-in mice were generated using a CRISPR/Cas9 based approach. Briefly, two sgRNAs were designed by CRISPR design tool (http://www.sanger.ac.uk/) to target either a region upstream of the exon 2 and downstream of the 3’UTR and then were screened for on-target activity using a Universal CRISPR Activity Assay. T7 promoter sequence was added to the Cas9 or sgRNA template by PCR amplification in vitro. Different concentrations of donor vector and purified in vitro transcribed Cas9 mRNA and sgRNAs were mixed and co-injected into the cytoplasm of one-cell stage fertilized eggs of C57BL/6N mice. The injected zygotes were transferred into oviducts of pesudopregnant females to generate F0 mice. PCR and sequencing verification of founder pups harboring the intended mutation were then crossed with wild-type mice for germline transmission, which were further confirmed by PCR, sequencing and Southern blot.
Sundby LJ, Hawbaker KM, Powers J, Southern WM, Johnson EE, Patrinostro X, Perrin BJ, Ervasti JM. The complete absence of cytoplasmic γ-actin results in no discernible phenotype in mice or primary fibroblasts. FEBS J. 2025 Mar 20. doi: 10.1111/febs.70075. Epub ahead of print. (Medline PMID: 40109120)
Colony Surveillance Program and Current Health Reports
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@missouri.edu 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 Naomi Walsh.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Naomi Walsh
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.
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.
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