Strain Detail Sheet

Strain Name:
C57BL/6N-Chrnb2tm1.1Geno/Mmucd
Stock Number:
066430-UCD
Citation ID:
RRID:MMRRC_066430-UCD
Other Names:
C57BL/6ChrnBeta2-GFP

Strain Information

Gene Details

Chrnb2tm1.1Geno
Name: cholinergic receptor nicotinic beta 2 subunit; targeted mutation 1.1, Genoway
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 3
Alteration at locus: Knock-In
Chrnb2
Name: cholinergic receptor nicotinic beta 2 subunit
Synonyms: [b]2-nAchR, Acrb-2, Acrb2, C030030P04Rik
Type: Gene
Species: Mouse
Chromosome: 3
Alteration at locus: Knock-In
NCBI: 11444
HGNC: HGNC:1962
Homologene: 595
Genome Browser [Click image to go to Jbrowse]



Additional Resources
IMPC
Google
Entrez
PubMed
IMSR
Gene Ontology
BioGPS
Gene Cloud
IGTC
Mouse Phenome DB
Mouse Mine
KOMP Phenotyping
Monarch
GeneCards
ClinGen
Genetic Alterations
In-frame insertion of monomeric GFP into exon 5 of the Chrnb2 gene, which encodes the M3–M4 intracellular loop of the nicotinic ACh receptor beta2 subunit. The targeting vector was also contained a neomycin selection cassette flanked by loxP sites, several kilobases of homologous 5′ and 3′ sequence (homology arms), and a diphtheria toxin-negative selection cassette.The neomycin cassette was removed via Cre deletion.
Genotype Determination
Genotyping Protocol
ES Cell Line
genOway ESC line derived from C57BL/6N

Strain Description

Phenotype
None/Normal/Wild-type
MeSH Terms
  • Animals
  • Cells, Cultured
  • Chloride Channels/analysis
  • Clone Cells
  • Dendrites/metabolism
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Mesencephalon/cytology
  • Mesencephalon/metabolism
  • Mice
  • Microscopy, Fluorescence
  • Neurons/chemistry
  • Neurons/drug effects
  • Neurons/metabolism
  • Nicotine/pharmacology
  • Patch-Clamp Techniques
  • Protein Subunits
  • Protein Transport
  • Receptors, Nicotinic/analysis
  • Receptors, Nicotinic/metabolism
  • Receptors, Nicotinic/physiology
  • Up-Regulation
  • Choline O-Acetyltransferase/metabolism
  • Dose-Response Relationship, Drug
  • Electric Stimulation
  • Excitatory Amino Acid Agents/pharmacology
  • Female
  • Gene Expression/drug effects
  • Gene Expression/genetics
  • Green Fluorescent Proteins/genetics
  • Habenula/cytology
  • Habenula/drug effects
  • Habenula/metabolism
  • In Vitro Techniques
  • Male
  • Membrane Potentials/drug effects
  • Membrane Potentials/genetics
  • Membrane Potentials/physiology
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Neurons/physiology
  • Protein Subunits/genetics
  • Protein Subunits/metabolism
  • Receptors, Nicotinic/genetics
  • alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid/pharmacology
Strain Development
A targeting vector was constructed containing exon 5 with GFP fused in-frame in the desired position in the M3-M4 loop coding sequence. Several kilobases of 5′ and 3′ sequences (relative to exon 5) were included to direct homologous recombination. A neomycin cassette flanked by loxP sites was inserted adjacent to the modified exon 5 in the targeting vector. A diphtheria toxin negative selection cassette was inserted adjacent to one of the homology arms (5′ or 3′). Targeting vectors were linearized and electroporated into C57BL/6N embryonic stem cells. G418-resistant cell clones were isolated and amplified.

First, clones were screened via PCR to identify those positive for both the 5′ and 3′ homologous recombination events. Next, clones positive in the 5′ and 3′ PCR assays were further evaluated for correct homologous recombination (and the absence of additional genomic rearrangements) via Southern blot with probes designed to detect both the 5′ and 3′ homologous recombination events. At least three positive ES cell clones were injected into 3.5-d-old embryos (blastocyst stage) derived from an albino C57BL/6 strain (B6(Cg)-Tyrc-2J/J - RRID:IMSR_JAX:000058), reimplanted into pseudo-pregnant females, and allowed to develop to term. Progeny males with evidence of high (>50%) chimerism (black vs. white coat color) were selected for further breeding. To breed chimeras to the F1 generation and to excise the neomycin selection cassette from the genome of the F1 progeny, chimeric males were mated with female proprietary to genOway Cre-deleter mice on C57BL/6N background. To confirm germline transmission, neo-excision, and maintenance of the proper homologous recombination event, F1 mice were genotyped via both PCR and Southern blot. The β2-GFP mice are fully isogenic with C57BL/6N.
Suggested Control Mice
C57BL/6N

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@ucdavis.edu. Older strains may not have this information.

Research Applications

  • Models for Human Disease
  • Neurobiology

Strain Origin

Donor
Henry Lester, Ph.D., California Institute of Technology.
Ryan Drenan, Ph.D., Wake Forest University, School of Medicine.
Primary Reference

Nashmi R, Dickinson ME, McKinney S, Jareb M, Labarca C, Fraser SE, Lester HA. Assembly of alpha4beta2 nicotinic acetylcholine receptors assessed withfunctional fluorescently labeled subunits: effects of localization, trafficking, and nicotine-induced upregulation in clonal mammalian cells and in culturedmidbrain neurons. J Neurosci. 2003 Dec 17;23(37):11554-67. (Medline PMID: 14684858)

Shih PY, Engle SE, Oh G, Deshpande P, Puskar NL, Lester HA, Drenan RM.Differential expression and function of nicotinic acetylcholine receptors insubdivisions of medial habenula. J Neurosci. 2014 Jul 16;34(29):9789-802. doi:10.1523/JNEUROSCI.0476-14.2014. (Medline PMID: 25031416)

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@ucdavis.edu.

Appearance

Coat Color
Black
Eye
Black

Breeding

MMRRC Breeding System
Random intra-strain mating
Generation
N6+ (C57BL/6N)
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: 4 to 6 months 10 to 12 months
Average litter size
4 to 6
Recommended wean age
3 Weeks
Average Pups Weaned
4 to 6

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@ucdavis.edu for more information.

Conditions of Distribution

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
066430-UCD-SPERM Cryo-preserved spermatozoa $546.25 / $869.88
Non-Profit / For-Profit		 Aliquot Approximate quantity3
066430-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.