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Updated: Jun 10, 2025

Substrate Generation for Endonucleases of CRISPR/Cas Systems
Published on: September 8, 2012
Model-directed generation of artificial CRISPR-Cas13a guide RNA sequences improves nucleic acid detection
Sreekar Mantena1,2,3, Priya P Pillai1, Brittany A Petros1,4,5,6
1Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Abstract:
CRISPR guide RNA sequences deriving exactly from natural sequences may not perform optimally in every application. Here we implement and evaluate algorithms for designing maximally fit, artificial CRISPR-Cas13a guides with multiple mismatches to natural sequences that are tailored for diagnostic applications. These guides offer more sensitive detection of diverse pathogens and discrimination of pathogen variants compared with guides derived directly from natural sequences and illuminate design principles that broaden Cas13a targeting.
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