MTA required. The Gad67-CreERT2 rat model was generated using CRISPR/Cas9 for use as a breeding mate for conditional knockout/ knock-in rat models. To achieve this, a pair of in vivo validated gRNAs, a single-stranded deoxyribonucleic acid (ssDNA) donor, and the Cas9 protein were injected into the mouse embryos. Mice born from the injection were screened by genotyping PCR and NGS/ Sanger sequencing to confirm founders (F0). The Cre rat line generated expresses Cre recombinase under the control of tissue-specific gene promoter Gad67. Furthermore, DNA recombinase activity is induced by Tamoxifen, so that the Cre-Lox model can be both conditional and inducible.
Tissue/Cell-specificity |
Rat line |
Promoter |
Reference |
GABAergic neurons, islet cells and spermatocytes |
GAD67-CreERT2
|
10.2-kb containing 8.4-kb promoter and noncoding exon 0A and 0B of mouse glutamate decarboxylase 67 |
Kabayashi et al 2003 Rasmussen et al 2007 |