Table 7.
Summary of Physical Transfection Techniques for CRISPR Delivery
| Transfection Method | Cell Type | CRISPR Format & Dosage | Gene Editing Performance | Target Gene | Ref. |
|---|---|---|---|---|---|
| Mechanical | SU-DHL-1 lymphoma cells | Plasmid , @ - | 70% knockout of fluorescence expression, assessed via flow cytometry | EGFP | 88 |
| MDA-MB-231 basal cells | Plasmid , @ - | 90% knockout of fluorescence expression, assessed via flow cytometry | EGFP | ||
| SK‐BR‐3 cells | RNP, @ 2 µM | 80% knockout of fluorescence expression, assessed via flow cytometry | EGFP | 90 | |
| MDA-MB-231 basal cells | RNP, @ 2 µM | 43% editing, assessed by surveyor mutation detection assay | pMAPKs | ||
| SUM-159 cells | RNP, @ 2 µM | 47% editing, assessed by surveyor mutation detection assay | pMAPKs | ||
| Human primary T cells | RNP, @ 2 µM | 36% editing, assessed by surveyor mutation detection assay | pMAPKs | ||
| ~35% knockout of PD-1 surface expression, assessed via flow cytometry | PD-1 | ||||
| Human HSCs | RNP, @ 2 µM | - | C/EBPα | 87 | |
| Human HSCs | RNP, @ 25 µM | 63% knockout of β2-microglobulin surface expression, assessed via flow cytometry | B2M | 89 | |
| Human primary T cells | RNP, @ 0.625 µM | 55% editing, assessed by T7E1 assay | PD1 | 91 | |
| Electrical | HEK 293 cells | Plasmid, @ 200 µg/mL | - | - | 92 |
| Human primary CD4+ T cells | RNP @ ~ 50 µM | 96% knockout of CXCR4 surface expression, assessed via flow cytometry | CXCR4 | 110 | |
| 98% knockout of CD127 surface expression, assessed via flow cytometry | CD127 | ||||
| 94% knockout of CCR7 surface expression, assessed via flow cytometry | CCR7 | ||||
| Human primary CD8+ T cells | 95% knockout of PD1 surface expression, assessed via flow cytometry | PD1 | |||
| 96% knockout of TIGIT surface expression, assessed via flow cytometry | TIGIT | ||||
| 98% knockout of CTLA4 surface expression, assessed via flow cytometry | CTLA4 | ||||
| Mouse CD4+ T cells | 90% knockout of CD90 surface expression, assessed via flow cytometry | CD90 | |||
| 88% knockout of CTLA4 surface expression, assessed via flow cytometry | CTLA4 | ||||
| Mouse CD8+ T cells | 98% knockout of CD8α surface expression, assessed via flow cytometry | CD8α | |||
| 93% knockout of CTLA4 surface expression, assessed via flow cytometry | CTLA4 | ||||
| Human iPSCs | RNP, @ 0.5 µM | 52% editing of APP, assessed by T7E1 assay | APP | 94 | |
| 70% editing of AAVS1, assessed by T7E1 assay | AAVS1 | ||||
| 59% editing of OCT4, assessed by T7E1 assay | OCT4 | ||||
| 52% editing of PD1, assessed by T7E1 assay | PD1 | ||||
| 56% editing of APP, assessed by DNA sequencing90% editing of APP with HDR DNA template, assessed by DNA sequencing48% editing of PD1, assessed by DNA sequencing62% editing of PD1 with HDR DNA template. assessed by DNA sequencing | |||||
| Human MSCs | RNP, @ 0.5 µM | 45% editing of B2M, assessed by T7E1 assay30% knockout of β2-microglobulin surface expression, assessed via flow cytometry77% knockout of β2-microglobulin surface expression with HDR DNA template, assessed via flow cytometry | B2M | ||
| Human primary T cells | RNP, @ 0.5 µM | 57% editing of PD1, assessed by T7E1 assay14% knockout of PD1 surface expression, assessed via flow cytometry19% knockout of PD1 surface expression with HDR DNA template, assessed via flow cytometry | PD-1 | ||
| HEK 293 cells | RNP, @ 10 µM | 31% editing, assessed by T7E1 assay33% editing, assessed by Sanger sequencing | PPIB | 95 | |
| HeLa cells | RNP, @ 10 µM | 24% editing, assessed by T7E1 assay | PPIB | 101 | |
| Jurkat cells | RNP, @ 10 µM | 26% editing, assessed by T7E1 assay | |||
| Acoustic | HEC-1A cells | Plasmid, @ 2 μg/mL | 57% knockout of mRNA expression, assessed via western blot | C-erbB-2 | 118 |
| HeLa & HEK 293 cells | Plasmid, @ 110 ng/μl | - | - | 121 | |
| Magnetic | Porcine fibroblast | Plasmid, @ 375 ng/mL & 750 ng/mL | 15% & 27% editing, assessed by Sanger sequencing | H11 locus | 127 |
| Hepa 1-6 cell | Plasmid in baculovirus | 35% editing, assessed by T7E1 assay | Vegfr2 | 129 | |
| Laser/ Plasmonic | SC1 cells | sgRNA (to Cas9 expressing cells), @ 5 µM | 12% editing, assessed by T7E1 assay30% knockout of CCR7 surface expression, assessed via flow cytometry | CCR7 | 141 |
| RNP, @ 5 µM | 6% editing, assessed by T7E1 assay22% knockout of CCR7 surface expression, assessed via flow cytometry | ||||
| Mouse primary T cells | RNP, @ 5 µM | 4% knockout of CXCR3 surface expression, assessed via flow cytometry | CXCR3 | ||
| Lymph node stroma cells | RNP, @ 5 µM | 5% knockout of fluorescence expression, assessed via flow cytometry | EGFP |