A Biophysical Model of CRISPR/Cas9 Activity for Rational Design of Genome Editing and Gene Regulation
(A) The addition of target DNA sites with the same sequence sequesters the Cas9:crRNA complex, and increases the transcription rate of the promoter controlling YFP expression. (B) A comparison between model-calculated transcription rates and measured YFP expression levels when either (stars) 0, (circles) 1, (diamonds) 2, (squares) 4, or (triangles) 8 additional on-target DNA sites were added. The DNA sites’ initial superhelical densities were either (left) increased by 0.0065 per occupied site or (right) kept constant. Data points and bars represent the mean and standard deviation of 2 measurements, performed in this study.