Table 6.

Comparison of Current Physical Transfection Methods for CRISPR/Cas9

Physical Methods	Advantages	Drawbacks	Suitable cargo	Recommended Cell Condition
Mechanical (non-microinjection)	Effective to difficult-to-transfect cells Simple setup and infrastructure Do not require external field supply	Ineffective delivery for large molecules	Protein mRNA	Suspension
Electrical	Effective to difficult-to-transfect cells Applicable for in situ transfection	May damage cells and cargo molecules	pDNA mRNA	Suspension (bulk) Adherent (micro/nano)
Acoustoporation	High cell viability	Limited throughput Low efficiency	Protein mRNA	Adherent
Laser / optothermal	High spatial control	Low throughput May damage cells at certain wavelengths	Protein mRNA	Adherent
Magnetic	Effective to difficult-to-transfect cells Applicable for in situ transfection	Require chemical complex formation	pDNA mRNA	Adherent