| The Type II prokaryotic CRISPR/Cas system is the new class of tools for targeted genome engineering. The cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-cas systems uses small RNAs as guides (gRNA) to cleave DNA in a sequence-specific manner. With its ease in designing guide sequences to target specific genomic loci, the CRISPR/Cas system is a much simpler, faster, and robust alternative to TALEN and Zinc finger nuclease platforms. For site-specific genome editing, the CRISPR/Cas9 system requires the Cas 9 nuclease and the gRNA. This RGD-fiber modified adenovirus expresses a codon-optimized Cas9 nuclease with N-terminal FLAG tag and a mCherry reporter. It can be used along with guided RNA for genome-editing in cells that regular Ad5 doesnt infect well, including human or mouse macrophages, ES cells, T cells, synoviocytes, certain fibroblast and islet grafts etc. |
