FVB.B6N-Gnb1^em2Frk/DgoldJ

Stock number: 036596
Product name: Gnb1^D76G
Research areas: Cancer Research, Cell Biology Research

Description

Gnb1^D76G knockin mice express the D76G (aspartic acid to glycine) amino acid substitution in the Gnb1 (guanine nucleotide binding protein (G protein), beta 1) locus. The mutation results in homozygous perinatal or embryonic lethality. The mutation is associated with neurodevelopmental disease and lymphocytic abnormalities.

Detailed description

Gnb1 (guanine nucleotide binding protein (G protein), beta 1) encodes the gamma subunit of membrane bound GTPases heterotrimeric G proteins. G protein regulate signals between receptors and effector proteins are involved in multiple signaling pathways including the PI3K/AKT/mTOR and RAS/MAPK pathways. . Mutations in GNB1 (including D76G) are associated with severe neurodevelopmental disease (PMID 27108799, 30194818, 32134617) and may also include lymphocytic abnormalities.
Gnb1^D76G knock-in mice were created using CRISPR/cas9 genome editing to express the D76G (aspartic acid to glycine) amino acid (aa) substitution in the Gnb1 locus. Heterozygous mice exhibit a small size, tail necrosis and some perinatal lethality. In addition, heterozygotes have a low electroconvulsive threshold when examined on the (B6 x FVB/NJ)F1 hybrid background. . The mutation results in homozygous perinatal or embryonic lethality. Additional characterization of this strain may be included at a later date. (July 2021)

This allele is also available on the C57BL/6N background, C57BL/6NJ-Gnb1^em2Frk/DgoldJ (Stock No. 030131).

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype could vary from that originally described.

Development

The Gnb1^D76G knock-in allele was created through the JAX Center for Precision Genomics (JCPG) at the The Jackson Laboratory. CRISPR/cas9 genome editing was used to create an amino acid substitution at position 76 (aspartic acid to glycine, GAT to GGC, D76G) as well as an unintended silent mutation, S74S (TCT to TCC), in the Gnb1 locus on chromosome 4. Single guide RNA and Cas9 endonuclease were introduced into C57BL/6NJ-derived fertilized eggs by pronuclear injection. Embryos were transferred to pseudopregnant females, and correctly targeted pups were bred to C57BL/6NJ for germline transmission. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6NJ (Stock No. 005304) to develop the colony. Dr. David Goldstein and Dr. Wayne Frankel transferred the mice to Columbia University where they were backcrossed to FVB/N for at least 6 generations, and then sent back to The Jackson Laboratory as Stock No. 036596. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize FVB/NJ (Stock No. 001800) oocytes.

Allele

Symbol: Gnb1^em2Frk
Name: endonuclease-mediated mutation 2, Wayne N Frankel
MGI accession ID: MGI:7266713
Generation method: Endonuclease-mediated
Attributes: Not Specified
Synonyms: Gnb1^D76G
Marker symbol: Gnb1
Marker name: guanine nucleotide binding protein (G protein), beta 1
Marker synonyms: HG2A; MDS; MRD42
Chromosome: 4
Strain of origin: C57BL/6NJ
Molecular note: CRISPR/cas9 genome editing is used to create an amino acid substitution at position 76 (aspartic acid to glycine, GAT to GGC, D76G) as well as an unintended silent mutation, S74S (TCT to TCC).