CRISPR/cas9 endonuclease mediated genome editing of the Plcg2, phospholipase C, gamma 2, gene was used to introduce a 13 bp knock-out mutation (nucleotides 1570 to 1582). The targeted Plcg2 gene encodes a transmembrane enzyme that acts as a signaling molecule in such diverse biological processes as hematopoietic cells differentiation, tissue morphogenesis, autoinflammation, antibody deficiency, and autoimmunity. Mutations in this gene have been associated with PLCG2 associated antibody deficiency and immune dysregulation (PLAID). As the mice are characterized, we will modify the strain description and add phenotype data. Homozygous viability/fertility has not been tested (June 2018).
Signal guide RNA and donor oligonucleotide designed to introduce a knock-out mutation in the Plcg2 gene in which 13 bp are deleted (nucleotides 1570 to 1582) and the cas9 nuclease were introduced into the cytoplasm of B6(SJL)-Apoetm1.1(APOE*4)Adiuj/J (Stock#27894) derived fertilized eggs with well recognized pronuclei. Correctly targeted embryos were transferred to pseudopregnant females. Correctly targeted pups were identified by sequencing and PCR and further bred to C57BL/6J (Stock No. 000664) to breed out the Apoetm1.1(APOE*4)Adiuj allele and develop the colony.
|Allele Name||endonuclease-mediated mutation 2, MODEL-AD Center|
|Allele Type||Endonuclease-mediated (Null/Knockout)|
|Gene Symbol and Name||Plcg2, phospholipase C, gamma 2|
|Strain of Origin||B6(SJL)-Apoetm1.1(APOE*4)Adiuj/J|
|Molecular Note||CRISPR/cas9 endonuclease mediated genome editing is used to delete 13 bp (nucleotides 1570 to 1582) creating a null allele.|
Heterozygotes may be bred to C57BL/6J (Stock No. 000664) or wildtype littermates. As the mice are characterized, we will modify the strain description if necessary and add data related to viability and fertility. Homozygous viability/fertility has not been tested (June 2018).
When using the Plcg2 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #029910 in your Materials and Methods section.