Back

TOP-SECRETS enables Cas9 nucleases to discriminate SNVs outside of PAMs

Herring-Nicholas, A.; Fisher-Huynh, S.; Josephs, E. A.

2025-05-10 bioengineering
10.1101/2025.05.06.652491 bioRxiv
Show abstract

RNA-guided CRISPR nuclease Cas9 cannot reliably differentiate between single nucleotide variations (SNVs) of targeted DNA sequences determined by their guide RNA (gRNA): they typically exhibit similar nuclease activities at any of those variations, unless the variation occurs with specific sequence contexts known as protospacer adjacent motifs (PAMs). Our approach, "TOP-SECRETS," generates gRNA variants that allow Cas9 ribonucleoproteins (RNPs) to reliably discriminate between healthy and disease-associated SNVs outside of PAMs.

Matching journals

The top 3 journals account for 50% of the predicted probability mass.

50% of probability mass above

"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.