mRNA-mediated delivery of gene editing tools to human primary muscle stem cells

Cas9-induced single cut enables highly efficient and template-free repair of a muscular dystrophy causing founder mutation

“…As we have shown before, nucleofection of mRNA provides a gentle and efficient delivery platform with optimal nucleofection efficiency resulting in homogeneous transgene expression in PHSats. 49 Compared with plasmid-based delivery, mRNA strongly increased the editing efficiency and cell survival. Furthermore, mRNA delivery eliminates the risk of unwanted integrations of dsDNA into the host genome and can be directly translated in the cytoplasm without the need of entering the nucleus for transcription which is favored only during mitotic breakdown and reformation of the nuclear envelope.…”

Liver-specific in vivo base editing of Angptl3 via AAV delivery efficiently lowers blood lipid levels in mice

“…Further efforts would be required to develop novel approaches to offer temporal control of the gene editing reagents. Alternatively, transient delivery of base editors in the forms of ribonucleoprotein (RNP) [41][42][43] or mRNA 18,44,45 instead of DNA using non-viral approaches can bypass the size limitation, maintain the high editing efficiency and lower the OT activities. Lipid nanoparticle (LNP), formed by lipids and polyethylene glycol (PEG) encapsulating transgene materials, has been developed as a leading non-viral delivery method in gene therapy.…”

Cell-derived extracellular vesicles for CRISPR/Cas9 delivery: engineering strategies for cargo packaging and loading

“…For example, one can use external stimuli, such as light, to modulate the responses of EVs. Finally, CRISPR/Cas9 tools in the mRNA format 64,65 can also be packaged within EVs/exosomes as a means of delivering CRISPR/Cas9 tools, but only a few examples can be found in the literature. 47,66 Taken together, notwithstanding that clinical applications of EV-mediated genome therapy still face multiple technical hurdles, a bright future awaits ahead.…”