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Clustered Regularly Interspaced Palindromic Repeats (CRISPR) were discovered within bacteria as a self-defense mechanism against invading viruses by using CRISPR associated (Cas) proteins to cleave viral double-stranded DNA (dsDNA). Since then the CRISPR-Cas9 system has rapidly evolved into one of the most efficient and effective means of genome editing. Utilizing a single-guide RNA complementary to the target genome DNA sequence, the Cas9 enzyme from Streptococcus Pyogenes (SpyCas9) is able to recognize and cleave dsDNA. The goal of this research is to develop an in-vitro cleavage assay for the CRISPR-SpyCas9 system for further studies into the effects of RNA binding drugs such as Cisplatin and metal complexes on the cutting efficiency of the sgRNA-SpyCas9 complex.
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Fallah, Adam Ahmed, "Development of a CRISPR-SpyCas9 In-Vitro Cleavage Assay" (2020). Senior Projects Spring 2020. 295.
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