Strain Name:
B6J.B6N(SJL)-Hcrtr1tm1.1Ava/Mmnc
Stock Number:
065595-UNC
Citation ID:
RRID:MMRRC_065595-UNC
Other Names:
Hcrtr1 (floxed allele) MGI:5637400 or Hcrtr1-flox

Gene Information

Hcrtr1tm1.1Ava
Name: hypocretin (orexin) receptor 1; targeted mutation 1.1, Anne Vassalli
Synonyms: Hcrtr1flox
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 4
Alteration at locus: Conditional
GFP
Name: green fluorescent protein
Type: Gene
Species: Aequorea victoria (jellyfish)
Alteration at locus: Conditional
Hcrtr1
Name: hypocretin (orexin) receptor 1
Synonyms: OX1R
Type: Gene
Species: Mus musculus (mouse)
Chromosome: 4
Alteration at locus: Conditional
NCBI: 230777
HGNC: HGNC:4848
Homologene: 37492
Genetic Alterations
Conditional (floxed) Hcrtr1 knockout.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
  • ES Cell Line
    Ingenious Line, iTL IC1
    Phenotype
    This mouse line harbors a Cre-dependent, conditional KO allele of Hcrtr1, the gene encoding hypocretin/orexin neuropeptides' receptor type-1.

    Conditional phenotype: When crossed with a Cre-expressing line, expression of Hcrtr1 is inactivated and replaced with expression of GFP under the endogenous Hcrtr1 gene promoter. Selective inactivation of Hcrtr1 in noradrenergic or dopaminergic neurons impacts the mouse EEG in a number of ways during wakefulness as well as sleep, and to maladaptive behavior in various environmental contexts.

    MeSH Terms
    • Antibodies, Viral/immunology
    • Cross Reactions/immunology
    • Humans
    • Orexin Receptors/immunology
    • RNA-Binding Proteins/immunology
    • Viral Core Proteins/immunology
    • Adrenergic Neurons/physiology
    • Animals
    • Behavior, Animal
    • Brain/physiology
    • Gamma Rhythm
    • Locomotion
    • Mice
    • Mice, Knockout
    • Nesting Behavior
    • Orexin Receptors/deficiency
    • Orexins/metabolism
    • Theta Rhythm
    • Antigens, Ly/metabolism
    • Atherosclerosis/metabolism
    • Atherosclerosis/pathology
    • Atherosclerosis/prevention & control
    • Bone Marrow Cells/metabolism
    • Female
    • Hematopoiesis/drug effects
    • Hematopoiesis/physiology
    • Hypothalamic Area, Lateral/metabolism
    • Macrophage Colony-Stimulating Factor/biosynthesis
    • Macrophage Colony-Stimulating Factor/deficiency
    • Macrophage Colony-Stimulating Factor/metabolism
    • Male
    • Monocytes/drug effects
    • Monocytes/metabolism
    • Myelopoiesis/drug effects
    • Neutrophils/metabolism
    • Orexin Receptors/metabolism
    • Orexins/biosynthesis
    • Orexins/deficiency
    • Orexins/pharmacology
    • Sleep/drug effects
    • Sleep/physiology
    • Sleep Deprivation/metabolism
    • Sleep Deprivation/physiopathology
    • Sleep Deprivation/prevention & control
    Strain Development
    The targeting vector for homologous recombination was built using PAC RP23-12J1 (BACPAC Resources Center, Oakland, CA, USA) a PAC from a C57BL6/J genomic DNA library containing Hcrtr1. Homologous arms spanning in total 10.0 kb were synthesized by PCR amplification of PAC RP23-12J1 using LA Taq (Takara), and Accuprime High-Fidelity (Invitrogen) DNA polymerases. The targeting vector was linearized and electroporated in iTL IC1, an embryonic stem (ES) cell line derived from C57BL/6NTac mice (inGenious Targeting Laboratory, Ronkonkoma, NY, USA). Single colonies were screened by Southern blot analysis using probes external to the targeting vector in both 5′ and 3′, and a GFP internal probe. Two ES cell clones having undergone correct recombination events were identified and injected into BALB/cAnNHEW blastocysts. Resulting chimeras were mated to B6;SJL-Tg(ACTFLPe)9205Dym/J (RRID:IMSR_JAX:003800) transgenic mice to excise the FRT-neo-FRT cassette. The neo-excised final conditional KO allele was Hcrtr1tm1.1Ava (MGI: 5637400, Hcrtr1flox). The Hcrtr1flox mouse colony was expanded and maintained by intercrosses, crosses to various Cre lines (themselves in C57BL/6J genetic background), and a number of backcrosses to C57BL/6J mice (exact number unknown, N5-12). The resulting genetic background is therefore congenic with contributions from C57BL/6J, C57BL/6N (minor SJL components).
    Suggested Control Mice
    The donor's mouse behavioral and electrophysiological phenotyping was based on pairwise comparisons between a conditional knockout (CKO) mouse group and a control mouse group (CTR). Several Cre-lox breeding schemes were possible, including (e.g., using Dat-IRES-Cre mice for dopamine neuron-selective knockout):

    (1) Hcrtr1+/flox ; Dat+/IRES-Cre x Hcrtr1+/flox; Dat+/IRES-Cre
    • CTR: Hcrtr1+/+; Dat+/IRES-Cre
    • cKO: Hcrtr1flox/flox; Dat+/IRES-Cre
    (2) Hcrtr1flox/flox ; Dat+/+ x Hcrtr1flox/flox; Dat+/IRES-Cre
    • CTR: Hcrtr1flox/flox
    • cKO: Hcrtr1flox/flox; Dat+/IRES-Cre
    • Immunology and Inflammation
    • Models for Human Disease
    • Neurobiology
    Donor
    Anne Vassalli, Ph.D., University of Lausanne.
    Primary Reference

    Vassalli A, Li S, Tafti M. Comment on "Antibodies to influenza nucleoprotein cross-react with human hypocretin receptor 2". Sci Transl Med. 2015 Nov 18;7(314):314le2. doi: 10.1126/scitranslmed.aad2353. (Medline PMID: 26582896)

    Li S, Franken P, Vassalli A. Bidirectional and context-dependent changes intheta and gamma oscillatory brain activity in noradrenergic cell-specificHypocretin/Orexin receptor 1-KO mice. Sci Rep. 2018 Oct 19;8(1):15474. doi:10.1038/s41598-018-33069-8. (Medline PMID: 30341359)

    McAlpine CS, Kiss MG, Rattik S, He S, Vassalli A, Valet C, Anzai A, Chan CT,Mindur JE, Kahles F, Poller WC, Frodermann V, Fenn AM, Gregory AF, Halle L,Iwamoto Y, Hoyer FF, Binder CJ, Libby P, Tafti M, Scammell TE, Nahrendorf M,Swirski FK. Sleep modulates haematopoiesis and protects against atherosclerosis. Nature. 2019 Feb;566(7744):383-387. doi: 10.1038/s41586-019-0948-2. Epub 2019 Feb13. (Medline PMID: 30760925)

    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_health@med.unc.edu .
    Coat Color
    Black
    Eye
    Black
    MMRRC Breeding System
    Backcross
    Generation
    N5+ (C57BL/6J)
    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: 10 to 12 months Greater than 12 months
    Average litter size
    7 to 9
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    7 to 9

    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@med.unc.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
    065595-UNC-RESUS Litter recovered from cryo-archive $2,829.00 / $4,696.00
    Non-Profit / For-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.