Protein-based therapy has conventional limitations because of its low efficiency and the difficulty of targeted delivery. To overcome this, we designed that lecithin-based liposome (NL) containing a Cas9/sgRNA ribonucleoprotein (Cas9-RNP) complex were linked to a microbubble (MB). Importantly, the bursting of the MB under acoustic pressure released the NL and enhanced their intracellular delivery owing to a sonoporation effect. We demonstrated that the Cas9-RNP within the NL used for SRD5A2 gene editing improved cell proliferation by blocking conversion of testosterone from dihydrotestosterone (DHT) which induces cell death. Interestingly, multiple treatments of the microbubble-nanoliposome (MN) exhibited higher efficacy than a single treatment. In summary, these results demonstrated that the ultrasound active MN system incorporating Cas9-RNP for hair loss therapy enhanced gene editing efficiency in human dermal papilla cells by sonoporation. Also, we expect that this MN system reduce side effects by provide opportunities to local delivery of Cas9-RNP to dermal papilla cells in hair follicles.
