nuclease-dead Cas9 (dCas9) interactions
First-pass extracted concept
nuclease-dead Cas9
Candidate: toolkit item1 source documents4 linked claims
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Aliases
dCas9
Evidence Snippets
Supporting Sources
Linked Claims
PAM-distal mismatches produce synergistic changes in dCas9 association kinetics when other gRNA-target mismatches are present.
Quoted textsource-backed
and synergistic changes in association kinetics when other gRNA-target mismatches are present
A massively parallel high-throughput sequencing flow cell method was used to comprehensively assess effects of combinatorial mismatches between gRNA and target nucleotides, including seed, PAM-distal, and PAM-disrupting changes, on dCas9-DNA interactions.
Quoted textsource-backed
Using a massively parallel method to quantify protein-DNA interactions on a high-throughput sequencing flow cell, we comprehensively assess the effects of combinatorial mismatches between guide RNA (gRNA) and target nucleotides, both in the seed and in more distal nucleotides, plus disruption of the protospacer adjacent motif (PAM).
Cas9 specificity is modulated by PAM-distal gRNA-DNA mismatches through effects on biophysical parameters that determine association and dissociation rates.
Quoted textsource-backed
Together, these observations support a model for Cas9 specificity wherein gRNA-DNA mismatches at PAM-distal bases modulate different biophysical parameters that determine association and dissociation rates.
PAM-distal mismatches can reverse dCas9 binding at long time scales.
Quoted textsource-backed
We report two consequences of PAM-distal mismatches: reversal of dCas9 binding at long time scales