Strain Detail Sheet

Strain Name:
B6.129P2-Dcaf1tm1.1Yxi/Mmnc
Stock Number:
066517-UNC
Citation ID:
RRID:MMRRC_066517-UNC
Other Names:
VprBP Conditional Knockout, Xiong UNC

Strain Information

Gene Details

Dcaf1tm1.1Yxi
Name: DDB1 and CUL4 associated factor 1; targeted mutation 1.1, Yue Xiong
Synonyms: VprBPflox
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 9
Alteration at locus: Knockout
Dcaf1
Name: DDB1 and CUL4 associated factor 1
Synonyms: Vprbp, B930007L02Rik
Type: Gene
Species: Mouse
Chromosome: 9
Alteration at locus: Knockout
NCBI: 321006
Homologene: 8805
Genome Browser [Click image to go to Jbrowse]



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Genetic Alterations
A 9.3-kb fragment of mouse genomic DNA, spanning from exon 5 to intron 8 of the Dcaf1 gene, was amplified by PCR from a mouse 129SvEv embryonic stem (ES) cell genomic DNA. LoxP sites were inserted at the 5′ and 3′ ends of a 2,358-bp genomic fragment containing exons 7 and 8 encoding 203 amino acid residues (Val172 to Ala374). The neomycing cassette was removed via transient expression of FLPe recombinase.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
    • ES Cell Line
    E14 derived from 129P2/OlaHsd

    Strain Description

    Phenotype
    Conditional phenotype: Knockout of Dcaf1 in mice caused embryonic death prior to embryonic day 7.5 (E7.5), G0-to-G1 progression and subsequent expansion of T cells, indicating that Dcaf1 is essential for mouse embryonic development. Conditional inactivation of Dcaf1 in mouse cells or depletion using RNA interference (RNAi) in human cells produced cell cycle defects. The donor found that Dcaf1 binds to chromatin during cell cycle progression from S through G2 and depletion of Dcaf1 reduced the rate of DNA replication and blocked S phase progression. Their genetic studies revealed a distinct phenotype of Dcaf1 deletion—selective elimination of proliferating, but not non-dividing quiescent cells—that is shared with Ddb1 deletion and associated with p53 activation.

    The donor also found that conditional deletion of Dcaf1 in mouse brain and lens resulted in a series of defects, including selective elimination of proliferation, increased apoptosis and DNA damage, neuronal and lens degeneration, brain hemorrhage, and perinatal lethality. This suggested essential function of Dcaf1 in the regulation of ribosomal biogenesis which provides novel molecular insight for the function of Dcaf1 in cell proliferation and embryonic development.
    MeSH Terms
    • Amino Acid Sequence
    • Animals
    • Carrier Proteins/physiology
    • Catalytic Domain
    • Chromatin/enzymology
    • DNA-Binding Proteins/genetics
    • DNA-Binding Proteins/metabolism
    • Dioxygenases/metabolism
    • Female
    • HEK293 Cells
    • Humans
    • Male
    • Mice, Knockout
    • Molecular Sequence Data
    • Mutation, Missense
    • Protein Binding
    • Protein Interaction Domains and Motifs
    • Protein-Serine-Threonine Kinases
    • Proto-Oncogene Proteins/genetics
    • Proto-Oncogene Proteins/metabolism
    • Ubiquitin-Protein Ligases
    • Ubiquitination
    • Carrier Proteins/genetics
    • Carrier Proteins/metabolism
    • Cell Cycle/physiology
    • Cells, Cultured
    • Chromatin/metabolism
    • Cullin Proteins/genetics
    • Cullin Proteins/metabolism
    • DNA Replication
    • Embryo, Mammalian/physiology
    • HIV-1/genetics
    • HIV-1/metabolism
    • Mice
    • Mice, Inbred C57BL
    • Mice, Transgenic
    • RNA Interference
    • Ubiquitin-Protein Ligases/genetics
    • Ubiquitin-Protein Ligases/metabolism
    • vpr Gene Products, Human Immunodeficiency Virus/genetics
    • vpr Gene Products, Human Immunodeficiency Virus/metabolism
    • B-Lymphocytes/cytology
    • Carrier Proteins/chemistry
    • Homeodomain Proteins/metabolism
    • Transgenes
    • V(D)J Recombination
    • Cell Survival/genetics
    • Cell Survival/physiology
    • Cellular Reprogramming/genetics
    • Dioxygenases/genetics
    • Fertility/genetics
    • Gene Silencing
    • Gonadal Dysgenesis/genetics
    • HeLa Cells
    • Mixed Function Oxygenases
    • Oocytes/physiology
    • Ovary/physiopathology
    Strain Development
    A 9.3-kb fragment of mouse genomic DNA, spanning from exon 5 to intron 8 of the Dcaf1 gene, was amplified by PCR from mouse 129SvEv embryonic stem (ES) cell genomic DNA and verified by DNA sequencing. LoxP sites were inserted at the 5′ and 3′ ends of a 2,358-bp genomic fragment containing exons 7 and 8 encoding 203 amino acid residues (Val172 to Ala374). To select for homologous recombination, a neomycin resistant gene (Neor), flanked by Flp recognition target sites, was inserted immediately upstream of the 5′ LoxP deletion site, and a thymidine kinase negative selection marker was inserted upstream of exon 5. Following homologous recombination in E14 ES cells (129P2/OlaHsd), a FLPe recombinase expression vector waw transiently transfected to delete the Neor cassette. The recombination event was confirmed by Southern blot analysis with probes against either exons 5 and 6 (probe 2) or exons 7 and 8 (probe 1). Three independent clones were injected into C57BL/6 blastocysts, and the resulting chimeras were mated with C57BL/6 females to generate Dcaf1floxΔ/+ heterozygous mice. Transmission of the targeted loci was confirmed by Southern blotting and PCR. Heterozygous offspring were intercrossed to produce homozygous mutant animals.
    Suggested Control Mice
    Wild-type C57BL/6J

    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@med.unc.edu. Older strains may not have this information.

    Research Applications

    • Apoptosis
    • Cancer
    • Cell Biology
    • Developmental Biology
    • Internal/Organ
    • Metabolism
    • Models for Human Disease
    • Neurobiology
    • Reproduction
    • Research Tools

    Strain Origin

    Donor
    Yue Xiong, Ph.D., UNC - School of Medicine.
    Primary Reference

    Nakagawa T, Lv L, Nakagawa M, Yu Y, Yu C, D'Alessio AC, Nakayama K, Fan HY,Chen X, Xiong Y. CRL4(VprBP) E3 ligase promotes monoubiquitylation and chromatin binding of TET dioxygenases. Mol Cell. 2015 Jan 22;57(2):247-260. doi:10.1016/j.molcel.2014.12.002. Epub 2014 Dec 31. Erratum in: Mol Cell. 2015 Sep17;59(6):1043. (Medline PMID: 25557551)

    McCall CM, Miliani de Marval PL, Chastain PD 2nd, Jackson SC, He YJ, Kotake Y,Cook JG, Xiong Y. Human immunodeficiency virus type 1 Vpr-binding protein VprBP, a WD40 protein associated with the DDB1-CUL4 E3 ubiquitin ligase, is essentialfor DNA replication and embryonic development. Mol Cell Biol. 2008Sep;28(18):5621-33. doi: 10.1128/MCB.00232-08. Epub 2008 Jul 7. (Medline PMID: 18606781)

    Kassmeier MD, Mondal K, Palmer VL, Raval P, Kumar S, Perry GA, Anderson DK,Ciborowski P, Jackson S, Xiong Y, Swanson PC. VprBP binds full-length RAG1 and isrequired for B-cell development and V(D)J recombination fidelity. EMBO J. 2012Feb 15;31(4):945-58. doi: 10.1038/emboj.2011.455. Epub 2011 Dec 13. (Medline PMID: 22157821)

    Yu C, Zhang YL, Pan WW, Li XM, Wang ZW, Ge ZJ, Zhou JJ, Cang Y, Tong C, SunQY, Fan HY. CRL4 complex regulates mammalian oocyte survival and reprogramming byactivation of TET proteins. Science. 2013 Dec 20;342(6165):1518-21. doi:10.1126/science.1244587. Erratum in: Science. 2014 May 2;344(6183):370. (Medline PMID: 24357321)

    Colony and Husbandry Information

    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_health@med.unc.edu.

    Appearance

    Coat Color
    Black
    Eye
    Black

    Breeding

    MMRRC Breeding System
    Backcross and random intra-strain mating
    Generation
    N10+ (C57BL/6J), F10+
    Overall Breeding Performance
    Good

    Reproductive Statistics

    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 7 to 9 months
    Bred to Homozygosity
    Yes
    Average litter size
    7 to 9
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    7 to 9

    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@med.unc.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
    066517-UNC-RESUS Litter recovered from cryo-archive $2,914.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
    Cryopreserved material may be available upon request, please inquire to mmrrc@med.unc.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.