MTA required. The Pomc-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 Pomc. 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 |
Neurons involved in the control of food intake (arcuate nucleus (hypothalamus) and solitary tract nucleus (hindbrain)) |
Pomc-CreERT2 |
13-kb of 5′ and 2 kb of 3′ mouse Pomc flanking sequences
|
Padilla et al 2011 Young et al 1998 |