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 language | English |
|---|---|
| Article number | gkaf016 |
| Number of pages | 14 |
| Journal | Nucleic Acids Research |
| Volume | 53 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 27 Jan 2025 |
Bibliographical note
Publisher Copyright:© 2025 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research.
Funding
This study was supported by a Career Development Fellowship grant from the Medical Research Council to C.R. (MR/Y009703/1) and research grants from the Kennedy\u2019s Disease Association to C.R. and from the Muscular Dystrophy Association/American Association of Neuromuscular & Electrodiagnostic Medicine Foundation (578222) to C.R. W.F.L. was supported by the Medical Research Council programme grant to M.J.A.W. (MR/N024850/1). A.J.T and A.G.G were supported by the Marie Curie Initial Training Network EScoDNA [FP7 project 317110], and Oxford University Press John Fell Fund. A.G.G was also supported by a La Caixa Foundation Fellowship (ID: LCF/BQ/EU16/11560044). Funding to pay the Open Access publication charges for this article was provided by the University of Oxford Article Processing Charge (APC). Author contributions : A.G.G., C.S., W.F.L., and J.R. con- ducted experiments and analyzed the data. O.G.J., P.V., S.E.A., and J.B. provided materials and assistance. A.G.G. and C.R. wrote the paper, with input from all authors. A.G.G., Y.A., A.J.T., M.J.W., and C.R. supervised the study. This study was supported by a Career Development Fel- lowship grant from the Medical Research Council to C.R. (MR/Y009703/1) and research grants from the Kennedy's Disease Association to C.R. and from the Muscular Dys-trophy Association/American Association of Neuromuscu- lar & Electrodiagnostic Medicine Foundation (578222) to C.R. W.F.L. was supported by the Medical Research Council programme grant to M.J.A.W. (MR/N024850/1). A.J.T and A.G.G were supported by the Marie Curie Ini- tial Training Network EScoDNA [FP7 project 317110], and Oxford University Press John Fell Fund. A.G.G was also supported by a La Caixa Foundation Fellowship (ID: LCF/BQ/EU16/11560044). Funding to pay the Open Access publication charges for this article was provided by the Uni- versity of Oxford Article Processing Charge (APC). Data availability The main data supporting the findings of this study are avail- able within this paper and its Supplementary Data. The raw and analyzed datasets are too large to readily share publicly, yet are available for research purposes from the corresponding author on reasonable request. References
| Funders | Funder number |
|---|---|
| Kennedy's Disease Association | |
| American Association of Neuromuscu- lar & Electrodiagnostic Medicine Foundation | |
| Muscular Dys-trophy Association | |
| Oxford University Press John Fell Fund | |
| Muscular Dystrophy Association | |
| University of Oxford | |
| Seventh Framework Programme | 317110 |
| Medical Research Council | MR/Y009703/1 |
| Fundación Bancaria Caixa d'Estalvis i Pensions de Barcelona | LCF/BQ/EU16/11560044 |
| American Association of Neuromuscular & Electrodiagnostic Medicine Foundation | MR/N024850/1, 578222 |
