Abstract
We present our work on how secondary structure formation in target sequences impacts the structure, conformational dynamics, and cleavage activity of CRISPR-Cas9. G-quadruplex (GQ) structures with varying stabilities were used as model secondary structures and were placed either in the target strand (TS) or non-target strand (NTS). Using single molecule FRET approach, we observed different conformational states and dynamics depending on the stability of the GQ and the position of PQS. We also observed variations in Cas9-mediated DNA cleavage activity depending on the stability of GQ and its location with respect to the target site.
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