Tissue-specific modulation of CRISPR activity by miRNA-sensing guide RNAs

Antonio Garcia-Guerra*, Chaitra Sathyaprakash, Olivier G. De Jong, Wooi F. Lim, Pieter Vader, Samir El Andaloussi, Jonathan Bath, Jesus Reine, Yoshitsugu Aoki, Andrew J. Turberfield, Matthew J.A. Wood, Carlo Rinaldi*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Nucleic acid nanostructures offer unique opportunities for biomedical applications due to their sequence-programmable structures and functions, which enable the design of complex responses to molecular cues. Control of the biological activity of therapeutic cargoes based on endogenous molecular signatures holds the potential to overcome major hurdles in translational research: cell specificity and off-target effects. Endogenous microRNAs (miRNAs) can be used to profile cell type and cell state, and are ideal inputs for RNA nanodevices. Here, we present CRISPR MiRAGE (miRNA-activated genome editing), a tool comprising a dynamic single-guide RNA that senses miRNA complexed with Argonaute proteins and controls downstream CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) activity based on the detected miRNA signature. We study the operation of the miRNA-sensing single-guide RNA and attain muscle-specific activation of gene editing through CRISPR MiRAGE in models of Duchenne muscular dystrophy. By enabling RNA-controlled gene editing activity, this technology creates opportunities to advance tissue-specific CRISPR treatments for human diseases.

Original languageEnglish
Article numbergkaf016
Number of pages14
JournalNucleic acids research
Volume53
Issue number2
DOIs
Publication statusPublished - 27 Jan 2025

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