Sprague Dawley rat 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.
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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 |