Gene-editing

You have a gene to edit. Should you use Cas9, Cas12a, or a Base Editor? This capstone post walks you through the entire workflow—from choosing the right molecular "scissors" for your target to designing and validating your final guides.

Choosing a guide RNA that cuts your target is only half the battle. You also need one that doesn’t cut anywhere else. This post breaks down the MIT and CFD scoring systems, how off-target algorithms actually work, and which tools are best for predicting Cas9 and Cas12a off-targets.

pegRNA design has five parameters that each affect prime editing efficiency. Here are the principles behind PBS length, RT template, nick distance, and secondary structure — then how to apply them in PrimeDesign.

A guide RNA score is only useful if your target base lands in the editing window. This post covers how to count protospacer positions, use BE-Designer for CBE and ABE guide design, manage bystander edits, and handle PAM constraints — with worked examples for both editor types.

A guide RNA score is only useful if you know what it measures. This post covers the sequence rules that determine Cas9 guide efficiency, how on-target scoring algorithms work and what they were trained on, off-target risk metrics, how to read CRISPOR output, and what changes when designing for Cas12a.