What question did this study set out to answer?

To develop a transient extracellular nanovesicle platform for packaging and delivering CRISPR-Cas9 ribonucleoproteins to achieve therapeutic exon skipping without viral vector-associated risks.

March 13, 2020SHILAP Revista de lepidopterologíaOpen Access

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

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Why the study?

Prolonged expression of CRISPR-Cas9 and gRNA from viral vectors can cause off-target mutagenesis and immunogenicity, necessitating a transient delivery system for therapeutic genome editing.

Does NanoMEDIC induce efficient genome editing and exon skipping in hard-to-transfect cells and in vivo models?

Population

Human iPS cells, neurons, myoblasts, DMD patient iPS cell-derived skeletal muscle cells, and mice

Comparison

NanoMEDIC ribonucleoprotein delivery system

Design

Preclinical development and in vitro/in vivo study

Authors

PGPeter GeeMaxCyte (United States)MLMandy Siu Yu LungKyoto University YOYuya OkuzakiNagoya University

Discussion

Member takes

Implication

May inform DMD exon-skipping strategies; leaves open human translation from animal data.

Key Points

To develop a transient extracellular nanovesicle platform for packaging and delivering CRISPR-Cas9 ribonucleoproteins to achieve therapeutic exon skipping without viral vector-associated risks.
Engineered extracellular nanovesicles (NanoMEDIC) using chemically induced dimerization to recruit Cas9 and viral packaging signals with self-cleaving riboswitches to package sgRNA.
Tested delivery and genome editing efficiency in human induced pluripotent stem cells, neurons, myoblasts, and DMD patient-derived skeletal muscle cells.
Evaluated in vivo exon skipping following a single intramuscular injection in luciferase reporter mice and mdx mouse models.
NanoMEDIC facilitated efficient genome editing in hard-to-transfect cell lineages, including human iPS cells, neurons, and myoblasts.
Exon skipping reached over 90% efficiency in skeletal muscle cells differentiated from DMD patient iPS cells.
A single intramuscular injection induced permanent genomic exon skipping in both luciferase reporter mice and dystrophic mdx mice.

Structured PICO

Does NanoMEDIC induce efficient genome editing and exon skipping in hard-to-transfect cells and in vivo models?

Population

Various hard-to-transfect cell types (human induced pluripotent stem cells, neurons, myoblasts), skeletal muscle cells derived from Duchenne muscular dystrophy (DMD) patient iPS cells, luciferase reporter mice, and mdx mice

Intervention

NanoMEDIC (extracellular nanovesicle-based ribonucleoprotein delivery system for CRISPR-Cas9 protein and sgRNA)

Outcome

Genome editing efficiency and exon skipping efficiencysurrogate

NanoMEDIC provides an efficient, transient delivery system for CRISPR-Cas9 genome editing, demonstrating high exon skipping efficiency in DMD models in vitro and in vivo.

Cite This Study

Gee et al. (2020) studied this question.

synapsesocial.com/papers/69dc19f3ce788f95bfb64ee1 https://doi.org/10.1038/s41467-020-14957-y

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Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Also Consider

Also Consider