Table 1.
Nanoparticle delivery of CRISPR/Cas9.
| Delivery approaches | NP formulation | Characterization | CRISPR/Cas9 cargo | Efficiency | Application | References |
|---|---|---|---|---|---|---|
| Lipid nanoparticle | Bioreducible lipid-like materials: cholesterol, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and C16-PEG2000-ceramide | Bioreducible | Cas9: sgRNA complexed | 70% (HEK293-GFP) | in vitro and in vivo | Wang et al., 2019 |
| Amino-ester-derived lipid like nanomaterials | Biodegradability and low toxicity | Cas9 mRNA and sgRNA | 41% (eGFP signal) | in vitro and in vivo | Zhang et al., 2017 | |
| Lipid-like nanomaterials: FTT lipids, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000) | High delivery efficiency and biodegradability | Cas9 mRNA and sgRNA | ~60% (in vivo base editing of PCSK9) | in vivo | Shi et al., 2020 | |
| Lipid nanoparticle: ionizable lipid (LP01), cholesterol, and DSPC | Cas9 mRNA and sgRNA | >97% [transthyretin (Ttr) in serum] | in vitro and in vivo | Finn et al., 2018 | ||
| Zwitterionic amino lipids: cholesterol:PEG-lipid | Permanent DNA editing | Cas9 mRNA and sgRNA | 95% (HeLa-Luc-Cas9) 1.5–3.5% (Hepatocytes) |
in vitro and in vivo | Miller et al., 2017 | |
| Polymer | Chitosan | High efficacy and non-cytotoxicity | Cas9 RNPs | 12.50% | in vitro | Qiao et al., 2019 |
| Carboxymethyl chitosan with AS1411 ligands | Dual-targeting delivery and high efficiency | pDNA | >90% (CDK11 protein) | in vitro | Liu et al., 2018 | |
| DNA nano-structure | DNA nanoclews | Stable assembly and multiple editing | RNP | 80% (EGFP in U2OS) | in vitro | Sun et al., 2015b |
| Branched DNA-based nanoplatform | Biocompatible | RNP | 40% (genetic cleavage) | in vitro and in vivo | Liu et al., 2019 | |
| MicroRNA-responsive DNA nanoclews | Stimuli-responsive release | RNP | 45% (EGFP in HeLa) | in vitro and in vivo | Shi et al., 2020 | |
| Inorganic nanomaterials | Cationic arginine gold nanoparticles (ArgNPs) | High delivery efficiency | RNP | ∼90% (delivery efficiency) 23–30% (gene editing efficiency) |
in vitro | Yang et al., 2011; Mout et al., 2017; Mout and Rotello, 2017 |
| CRISPR-Gold | Biocompatibility | RNP | 40–50% (mGluR5 protein and mRNA) | in vitro and in vivo | Lee et al., 2018 | |
| CRISPR-Gold | Low local immunogenicity, multiple uses | RNP | 5.40% (restoration of Duchenne muscular dystrophy) | in vivo | Lee et al., 2017 | |
| Exosome | Exosome-liposome hybrid | Efficiently encapsulate plasmid | pDNA | ∼40% (Runx2 mRNA) | in vivo | Lin et al., 2018 |
| VSV-G protein ectosomes: split GFP | Efficient delivery | Cas9 protein | _ | in vitro and in vivo | Zhang et al., 2020 | |
| Engineered exosome: GFP-GFP nanobody | Efficient delivery | Cas9 protein | _ | in vitro | Ye et al., 2020 | |
| Engineered exosome: CD9-HuR exosomes | Enhanced encapsulation | Cas9 mRNA | _ | in vitro and in vivo | Li et al., 2019 | |
| NanoMEDIC | Efficient delivery and high cleavage activity | Cas9 protein | 90% (exon skipping efficiencies) | in vitro and in vivo | Gee et al., 2020 | |
| Red blood cell (RBC)-derived EVs | Efficient loading and delivery | Cas9 mRNA | ~32% (gene silencing) ~18% (loading capacity Cas9 mRNA) |
in vitro and in vivo | Usman et al., 2018 |