Abstract
The use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 has moved from bench to bedside in less than 10years, realising the vision of correcting disease through genome editing. The accuracy and safety of this approach relies on the precise control of DNA damage and repair processes to achieve the desired editing outcomes. Strategies for modulating pathway choice for repairing CRISPR-mediated DNA double-strand breaks (DSBs) have advanced the genome editing field. However, the promise of correcting genetic diseases with CRISPR-Cas9 based therapies is restrained by a lack of insight into controlling desired editing outcomes in cells of different tissue origin. Here, we review recent developments and urge for a greater understanding of tissue specific DNA repair processes of CRISPR-induced DNA breaks. We propose that integrated mapping of tissue specific DNA repair processes will fundamentally empower the implementation of precise and safe genome editing therapies for a larger variety of diseases.
Highlights
Reviewed by: Michael Aregger, National Cancer Institute (NCI), United States Chris Richardson, University of California, Santa Barbara, United States
The successful implementation of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 technologies in a clinical setting relies on a deeper understanding of the DNA repair mechanisms and pathways responsible for genetic replacement outcomes, as well as the activity and accessibility of these pathways in specific cell types and tissues
The genome and transcriptome of target cells impact the effectiveness of genome editing approaches
Summary
Despite the error-prone nature of end-joining pathways, there is mounting evidence indicating that the pattern of DNA repair following a Cas9-induced DSB is not stochastic (van Overbeek et al, 2016; Shou et al, 2018) Based on this observation, several studies have systematically analysed how sequences flanking the DSB impact repair outcome, leading to the important conclusion that template-free Cas editing can be predicted and applied to achieve a specific outcome (Allen et al, 2018; Shen et al, 2018). Since NHEJ is the default pathway in human cells, its inhibition has been exploited to favour HDR This has been achieved through the use of small-molecules targeting LIG4 or DNA-PKcs (Robert et al, 2015; Riesenberg and Maricic, 2018), ubiquitinvariants targeting 53BP1 (Canny et al, 2017), expression of factors that displace 53BP1 from DSBs (Nambiar et al, 2019), or 53BP1 dominant negative forms (Paulsen et al, 2017; Figure 1C). SUCCESS OF CRISPR-BASED THERAPIES DEPENDS ON UNDERSTANDING TISSUE SPECIFIC DNA REPAIR
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