Development of CRISPR-Cas9 for in vivo medical applications
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Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR) and its CRISPR-associated (Cas) proteins have revolutionized genetic engineering. Since its discovery, it has supplanted previous methods of gene editing with its versatility and ease-of-use. Furthermore, the possibility of developing CRISPR-Cas9-based treatment plans for genetic therapy has piqued the interest of scientists. Much research has been done to evaluate its potential for use in medicine as well as the obstacles that prevent it from clinical applications. Although CRISPR-Cas9 can be effective at inducing double strand breaks in DNA, its overall template insertion efficiency is often not sufficient. By default, somatic cells repair damage to their DNA by undergoing nonhomologous end joining (NHEJ), which is not compatible with precise gene insertion. For a desired sequence to be inserted at the break site, homology directed repair (HDR) is required. Several methods of NHEJ inhibition and HDR stimulation have been evaluated and were effective at increasing HDR efficiency in vitro, but these results cannot be generalized for in vivo environments. Accordingly, results from CRISPR-Cas9 research have shown more promise for hematological diseases than for others. In order to address this disparity, additional work on its efficacy in vivo must be done. CRISPR-Cas9 has the potential to become the basis of future treatments for genetic disorders. Prior to this, treatments will need to account for the complex nature of in vivo processes. This thesis examines the current literature to gauge how far CRISPR-Cas9 technology is from use on human patients.