MTA required. The Tie2-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 Tie2. 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 |
|
Vascular endothelial cells including brain and retinal capillary |
Tie2-CreERT2 |
2.1-kb promoter +1.7kb intron 1 enhancer of mouse Tie2 (vascular endothelial-specific receptor tyrosine kinase) |
Schlaeger et al 1997 Ohtsuki et al 2005 |