Therapeutic In Vivo Gene Editing Achieved by a Hypercompact CRISPR-Cas12f1 System Delivered with All-in-One Adeno-Associated Virus.

CRISPR-based gene therapies are making remarkable strides toward the clinic. But the large size of most widely used Cas endonucleases including Cas9 and Cas12a restricts their efficient delivery by the adeno-associated virus (AAV) for in vivo gene editing. Being exceptionally small, the recently engineered type V-F CRISPR-Cas12f1 systems can overcome the cargo packaging bottleneck and present as strong candidates for therapeutic applications. In this study, the pairwise editing efficiencies of different engineered Cas12f1/sgRNA scaffold combinations are systemically screened and optimized, and the CasMINI_v3.1/ge4.1 system is identified as being able to significantly boost the gene editing activity. Moreover, packaged into single AAV vectors and delivered via subretinal injection, CasMINI_v3.1/ge4.1 achieves remarkably high in vivo editing efficiencies, over 70% in transduced retinal cells. Further, the efficacy of this Cas12f1 system-based gene therapy to treat retinitis pigmentosa in RhoP23H mice is demonstrated by the therapeutic benefits achieved including rescued visual function and structural preservation. And minimal bystander editing activity is detected. This work advances and expands the therapeutic potential of the miniature Cas12f1 system to support efficient and accurate in vivo gene therapy.

Scalp acupuncture therapy combined with exercise can improve the ability of stroke patients to participate in daily activities.

OBJECTIVE: To observe the improvement of stroke patients' ability to participate in daily activities by using Scalp acupuncture therapy combined with exercise. METHODS: According to random number table, patients were divided into a treatment group, which received rehabilitation training and scalp acupuncture to retain needles at different times, and a control group, which received rehabilitation training alone. The Barthel index was used to evaluate the activities of daily living before and after treatment. RESULTS: The scores of Barthel index of all groups were higher than those before treatment, and the scalp acupuncture for 2 h combined with rehabilitation training group had the highest score. There was significant difference compared with pre-treatment and other treatment groups. CONCLUSION: Scalp acupuncture therapy combined with exercise therapy can significantly improve the ability of stroke patients to participate in daily activities; 2 h of scalp acupuncture combined with exercise therapy had the most significant effect.