Harnessing novel CRISPR systems for genome engineering

RNA plays important and diverse roles in biology, but molecular tools to manipulate and measure RNA are limited. We demonstrate that RNA-targeting CRISPR effector Cas13 can be engineered for RNA diagnostics and mammalian cell RNA knockdown, binding, and editing. We demonstrate diagnostic applications of Cas13 for rapid and sensitive COVID-19 diagnostics. We also show Cas13 can be heterologously expressed in mammalian and plant cells for targeted knockdown of either reporter or endogenous transcripts, targeted RNA binding for transcript imaging, and programmable RNA editing. We develop engineered Cas13-based RNA editing tools for A-to-I and C-to-U base editing and show applications of these tools for therapeutics. Our results establish CRISPR-Cas13 as a flexible platform for RNA targeting with wide applicability for studying RNA or therapeutics.

Learning Objectives:

1. Learn about the discovery of Cas13 and its function as a programmable RNA-targeting CRISPR enzyme

2. Learn about how Cas13 can be engineered for nucleic acid diagnostics and for rapid, cheap, and sensitive COVID-19 detection

3. Learn about how Cas13 can be engineered for programmable RNA editing and therapeutic applications