The development of obesity and type 2 diabetes (T2D) is caused by a combination of genetic and lifestyle factors. Pharmacological options to sizably improve obesity and diabetes are scarce. The Crispr/Cas9 system has been repurpose to allow single or multiple target gene activation without editing the genome by fusion of a catalytically dead Cas9 (dCas9) to a transcriptional activation domain (VP64) [1, 2]. This transformative technology has been able to induce a physiologically relevant phenotype in a postnatal mammal and rescue genetic happloinsufficiencies diseases [3, 4]. In our group, we have been working with the production of a mouse model with the tissue specific expression of the Crispr/dCas9 activation system, and preliminary results showed multiple endogenous gene activation after a single adeno associated virus-gRNA injection. We hypothesize that the single or simultaneous co-activation of certain metabolic genes could be used to (1) reverse diet-induced obesity and to prevent diabetes progression in selected rodent models and (2) identify novel candidates genes that yield synergistic metabolic benefits.
Crispr/dCas-mediated rescue of genetic- and diet-induced obesity and type-2 diabetes
The aim of this project is to use the Crispr/dCas9 activation technology to selectively over activate either single target genes or simultaneously multiple target genes to improve body weight and glucose metabolism in selected mouse models of genetic- or diet-induced obesity. Analysis of gene function will include in vitro proof-of-principle in selected tissues and assessment of in vivo efficacy using state-of-the art technology. Successful PhD candidate will work in collaboration with Alberta Institute to analyze some of the biological samples produced during the experiments.
1. Gilbert L. et al. (2013). CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Cell 154, 2, 442-451.
2. Perez-Pinera P. et al. (2013). RNA-guided gene activation by CRISPR-Cas9– based transcription factors. Nature 10, 10, 973-976
3. Liao H. et al. (2017). In Vivo Target Gene Activation via CRISPR/Cas9-Mediated Trans-Epigenetic Modulation. Cell 171 (7) 1495-1507.
4. Matharu N. et al. (2019). CRISPR-mediated activation of a promoter or enhancer rescues obesity caused by haploinsufficiency. Science 363, 246.