Exosome–Liposome Hybrid Nanoparticles Deliver CRISPR/Cas9 System in MSCs

Abstract

Targeted delivery of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9) system to the receptor cells is essential for in vivo gene editing. Exosomes are intensively studied as a promising targeted drug delivery carrier recently, while limited by their low efficiency in encapsulating of large nucleic acids. Here, a kind of hybrid exosomes with liposomes is developed via simple incubation. Different from the original exosomes, the resultant hybrid nanoparticles efficiently encapsulate large plasmids, including the CRISPR–Cas9 expression vectors, similarly as the liposomes. Moreover, the resultant hybrid nanoparticles can be endocytosed by and express the encapsulated genes in the mesenchymal stem cells (MSCs), which cannot be transfected by the liposome alone. Taken together, the exosome–liposome hybrid nanoparticles can deliver CRISPR–Cas9 system in MSCs and thus be promising in in vivo gene manipulation.

Friend or Foe? Evidence Indicates Endogenous Exosomes Can Deliver Functional gRNA and Cas9 Protein

“…Several studies have used exosomes as vehicles to deliver the CRISPR/Cas9 expression plasmid into the host cells . However, it still needs to consider the potential off‐target effects which may be mediated by the large amount of gRNA and Cas9 protein constantly expressed from plasmid, and the integration risk of plasmid DNA itself.…”

Delivery of CRISPR/Cas9 by Novel Strategies for Gene Therapy

“…Researchers have developed exosome–liposome hybrid nanoparticles to deliver the CRISPR/Cas9 system into mesenchymal stem cells. The platform solves the challenges associated with a low loading capacity of the exosomes and difficulties in the transfection of mesenchymal stem cells with liposomes at the same time . Recently, on the basis of our work in the preparation of lipid‐based nanoparticles and gold nanoparticles, we developed a lipid/gold nanocluster delivery system that could successfully deliver Cas9 proteins and gPlk‐1 plasmids for gene therapy.…”

“…The platform solves the challenges associated with al ow loading capacity of the exosomes and difficulties in the transfection of mesenchymal stem cells with liposomes at the same time. [30] Recently,o nt he basis of our work in the preparationo fl ipidbased nanoparticles and gold nanoparticles, we developed a lipid/gold nanocluster delivery system that could successfully deliver Cas9 proteins and gPlk-1 plasmids for gene therapy.I n this delivery system,b esides effective encapsulation of the lipid shell, TAT-modified gold nanoclusters also condense the complex of Cas9 proteins and gRNA plasmids to decrease the packing volume and promote nuclear transport. After that, we createda nother photothermal-activatablel ipid/gold nanoparticle (AuNP) platform for the delivery of Cas9-gPlk-1 plasmids that could be thermally triggered to release TATp eptides from the AuNPs by cleavage of AuÀSb onds under l = 514 nm laser irradiation.…”

CRISPR/Cas9 Delivery Mediated with Hydroxyl‐Rich Nanosystems for Gene Editing in Aorta

“…Both viral and nonviral approaches are being evaluated for in vivo CRISPR/Cas9 delivery. [17,18] Some nonviral vectors, for example, liposomes, [19] polycations, [20] and inorganic nanoparticles, [9] were proved to be capable of delivering CRISPR/Cas9 system for gene editing. [16] In comparison, nonviral nanoparticles have the potential to overcome the limitations of large packaging capacity and serve as efficient and alternative DNA carriers due to their special structures, biocompatibility, and good membrane permeability.…”