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The Ctipco-rec allele lacks exon 2 of Ctip, including coding sequences for the initiator methionine M1 and the Ctip oligomerization domain. Ctipco-rec encodes an amino-terminally truncated CTIP protein that initiates at methionines M120 and/or M141 (Figure S2, PMID:32989150). The Ctipco-rec allele was derived from the conditional Ctipco allele, which harbors loxP sites flanking exon 2. Ctipco-rec/+ mice were originally generated by mating a heterozygous Ctipco/+ male with a female carrying the ubiquitously-expressed EIIa-cre transgene.
While heterozygous Ctipco-rec/+ mice develop normally and are fertile, homozygous Ctipco-rec/co-rec mice undergo early embryonic lethality.
The conditional-null Ctipco-neo targeting construct consists of a 7.7-kb genomic DNA fragment containing exon 1 and exon 2 of Ctip. The construct was generated by flanking exon 2, which encodes the N-terminal 36 amino acids of CTIP, with two loxP recombination signals. One loxP signal was introduced upstream of the transcriptional initiation site in intron 1, and the second loxP signal was inserted, along with a PGK-neomycin resistance cassette flanked by FRT (Flp recombinase target) signals, into intron 2. In addition, a herpes simplex virus type 1 thymidine kinase (HSV-TK) negative selection gene cassette was included to select against random integration. The conditional-null targeting construct was electroporated into 129/Sv embryonic stem (ES) cells and two independent neomycin-resistant ES clones harboring the Ctipco-neo knock-in allele were identified and injected into C57BL/6J blastocysts. Germline-transformed Ctipco-neo/+ heterozygous mice were mated with B6.Cg-Tg(ACTFLPe)9205Dym/J (RRID:IMSR_JAX:005703) mice to produce offspring in which the FRT-flanked neomycin cassette had been excised, thereby converting the Ctipco-neo allele into the desired Ctipco allele. The Ctipco/+ mice were backcrossed twice with C57BL/6J. The Ctipco-rec allele was then generated by mating Ctipco/+ mice with Rosa cre mice ubiquitously expressing cre recombinase (B6.FVB-Tg(EIIa-cre)C5379Lmgd/J - RRID:IMSR_JAX:003724). The Ctipco-rec/+ cre mice were intercrossed to obtain Ctipco-rec/+ mice.
Wang XS, Zhao J, Wu-Baer F, Shao Z, Lee BJ, Cupo OM, Rabadan R, Gautier J, Baer R, Zha S. CtIP-mediated DNA resection is dispensable for IgH class switch recombination by alternative end-joining. Proc Natl Acad Sci U S A. 2020 Oct 13;117(41):25700-25711. doi: 10.1073/pnas.2010972117. Epub 2020 Sep 28. (Medline PMID: 32989150)
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 Joan Jose Martinez.
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.
A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Joan Jose Martinez
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.