This Grin2bflox CRISPR/Cas9 generated mutant of the Grin2b gene possesses loxP sites flanking 599 nucleotides (exon 4 of Ensembl transcript 201 and exon 5 of transcript 202). This strain may be useful for generating conditional mutations in applications related to neurodevelopment.
Cathleen Lutz, The Jackson Laboratory
CRISPR/cas9 endonuclease mediated genome editing of the Grin2b, glutamate receptor, ionotropic, NMDA2B (epsilon 2), gene was used to introduce loxP sites flanking 599 nucleotides (exon 4 of Ensembl transcript 201 and exon 5 of transcript 202). The targeted Grin2b gene encodes part of the excitatory neurotransmitter N-methyl-D-aspartate (NMDA) receptor. Mutations in this gene have been associated with Mental Retardation, Autosomal Dominant 6, with or without Seizures and Early Infantile Epileptic Encephalopathy 27. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have loxP site flanked region deleted in the cre-expressing tissues. As the mice are characterized, we will modify the strain description and add phenotype data.
This model was generated in collaboration with the Simons Foundation Autism Research Initiative (SFARI).
Two guide RNAs were synthetized, a double stranded DNA donor plasmid containing loxP sequences flanking the second coding exon of Grin2b (exon 4 of Ensembl Grin2b transcript 201/ exon 5 of Ensembl transcript 202), and cas9 nuclease were co-injected into the cytoplasm of single cell C57BL/6J-derived zygotes containing well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by DNA sequencing of the targeted exon and flanking introns and further bred to C57BL/6J (Stock No. 000664) to develop the colony. Mice derived from founder 1087 (GET2481) were further bred to C57BL/6J inbred mice for two generations to develop the colony.
|Allele Name||endonuclease-mediated mutation 5, Cathy Lutz|
|Allele Type||Endonuclease-mediated (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Grin2b, glutamate receptor, ionotropic, NMDA2B (epsilon 2)|
|Strain of Origin||C57BL/6J|
|Molecular Note||CRISPR/Cas9 genome editing is used to insert loxP sequences flanking the second coding exon of Grin2b (exon 4 of Ensembl Grin2b transcript 201/ exon 5 of Ensembl transcript 202).|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Grin2b flox
mouse strain in a publication, please cite the originating article(s) and include JAX stock #032664 in your Materials and Methods section.