Effective delivery of genome editor to cervical cancer targeting Mcl1 for cancer therapy

CRISPR/Cas9 represents a transformative advancement in precision therapies, offering the promise of more effective and targeted treatment options. However, there are still limitations (including off-target editing as well as unsatisfied delivery tool) which obstruct the wide application of CRISPR/Cas9. Here, an endogenic artificial extracellular vesicles (EVs) system is engineered for effective delivery of Cas9 ribonucleoprotein (RNP). We demonstrated that the endogenic Cas9 RNP were sorted by the Lamp2b and delivered by the artificial EVs, which could markedly inhibit the growth of cervical cancer cells by inducing cell apoptosis. Moreover, artificial endogenic EVs RNP (Cas9-Mcl1) could result in remarkable antitumor effects in animal models of cervical cancer through suppressing Mcl1 expression. Our findings indicate that the artificial EVs delivery strategy could deliver Cas9 RNP effectively to inhibit cancer progression, which might be a promising treatment. Schematic illustration of Cas9 RNP delivery for targeting therapy. (A) The schematics for detecting the Mcl1 expression in cervical cancer from TCGA dataset and patients’ samples; (B) The recombinant “Lamp2b-PhoCl-Cas9” plasmid was constructed and transfected with HEK-293T. Lamp2b protein was expressed transmembrane of the EVs while Cas9 RNP were encapsulated into EVs; (C) The PhoCl protein could release Cas9 RNP freely into the EVs; (D) EVs RNP (Cas9-Mcl1) could deliver the (Cas9-Mcl1) RNP into the cervical cancer and suppress the cancer progression through editing the Mcl1 expression in cervical cancer.