Engineered RNA-Interacting CRISPR Guide RNAs for Genetic Sensing and Diagnostics

Galizi, Roberto; Duncan, John N.; Rostain, William; Quinn, Charlotte M; Storch, Marko; Kushwaha, Manish; Jaramillo, Alfonso

Abstract

CRISPR guide RNAs (gRNAs) can be programmed with relative ease to allow the genetic editing of nearly any DNA or RNA sequence. Here, we propose novel molecular architectures to achieve RNA-dependent modulation of CRISPR activity in response to specific RNA molecules. We designed and tested, in both living Escherichia coli cells and cell-free assays for rapid prototyping, cis-repressed RNA-interacting guide RNA (igRNA) that switch to their active state only upon interaction with small RNA fragments or long RNA transcripts, including pathogen-derived mRNAs of medical relevance such as the human immunodeficiency virus infectivity factor. The proposed CRISPR-igRNAs are fully customizable and easily adaptable to the majority if not all the available CRISPR-Cas variants to modulate a variety of genetic functions in response to specific cellular conditions, providing orthogonal activation and increased specificity. We thereby foresee a large scope of application for therapeutic, diagnostic, and biotech applications in both prokaryotic and eukaryotic systems.

Citation

Galizi, R., Duncan, J. N., Rostain, W., Quinn, C. M., Storch, M., Kushwaha, M., & Jaramillo, A. (2020). Engineered RNA-Interacting CRISPR Guide RNAs for Genetic Sensing and Diagnostics. The CRISPR Journal, 398 - 408. https://doi.org/10.1089/crispr.2020.0029

• Acceptance Date: Oct 20, 2020
• Publication Date: Oct 20, 2020
• Journal: The CRISPR Journal
• Print ISSN: 2573-1599
• Publisher: Mary Ann Liebert
• Pages: 398 - 408
• DOI: https://doi.org/10.1089/crispr.2020.0029
• Publisher URL: https://www.liebertpub.com/doi/pdf/10.1089/crispr.2020.0029