CRISPR/Cas9 has introduced new gene therapy possibilities for muscular dystrophies

Advancements in using CRISPR/Cas9 have introduced a host of new therapy possibilities for muscular dystrophies (MDs). There is a definite feeling of hope in the industry, but other barriers lay ahead, and they will define the future of MD gene editing. The ambiguity surrounding AAV transduction of satellite cells in vivo must be explained so that, if required, effort may be focused on optimizing vector targeting. Although the satellite cell correction needs are evident, it must be determined experimentally if high muscle turnover has a deleterious effect on CRISPR approaches. Another issue with muscular HDR is its low editing efficiency. Even outside the MD, exogenous, effective DNA integration would open up a slew of new possibilities.Either conventional HDR must be upgraded, or alternative techniques must be developed. The fact that both myotubes and latent satellite cells are post-mitotic means the latter are the most effective. Homology-independent targeted integration (HITI), homology-mediated end joining (HMEJ) and prime editing are three novel potentials. Duplication removal is another technique to restore full-length proteins. Duplications are the second most frequent DMD mutation, and a single sgRNA technique was used to restore dystrophin. To date, CRISPR/Cas9-mediated duplication removal has only been evaluated in DMD patient cells and must be tested in vivo. Because of their demonstrated track record in in vivo research and clinical trials, AAVs are expected to be employed in early generations of MD CRISPR therapy. Currently, AAVs may be the biggest choice, but future drugs will almost probably require a different delivery approach. It may take the shape of nanoparticles, which may carry a large range of transiently expressed payloads, while being very variable. If satellite cells can not be repaired, their capacity to escape immune reactions is crucial. To decrease the effects of muscle turnover, re-administration of nanoparticles may be utilized to treat MD throughout one's life. However, effective nanoparticle dosing for CRISPR in vivo editing has yet to be established in the muscle. Because this was not an AAV problem, the focus should be on new compositions of nanoparticles rather than improving the CRISPR/Cas9 system. The lack of published data suggests that nanoparticles' systemic muscle transport remains a considerable challenge. Due to muscle volume in the human body and the need to target muscles within the thoracic cavity, local intramuscular injections are not practical. Future research will focus primarily on developing an effective, muscle-specific nanoparticle that can be administered through circulation. The challenges ahead are tremendous, but with the appropriate focus and resources, answers will emerge, bringing therapeutic genome editing closer to the clinic than ever. While this research focused on DMD, the mentioned principles and methodology may and will undoubtedly be extended to several other MDs.