TABLE 5.

Summary of commonly used delivery methods for CRISPR/Cas9-mediated HBV gene editing.

Category	Methods	Features	Applications
In vitro delivery	Chemical Transfection	Chemical compounds facilitate the uptake of CRISPR/Cas9 complexes by cells	Simple and cost-effective for HBV gene editing in cell lines, with lower efficiency compared to other methods
	Electroporation	Brief electric pulses create transient pores in the cell membrane, allowing CRISPR/Cas9 entry	High transfection efficiency for difficult-to-transfect cells, suitable for HBV gene editing in primary hepatocytes and other challenging cell types
	Microinjection	CRISPR/Cas9 components are directly injected into individual cells using a fine needle	Precise delivery method suitable for HBV gene editing in primary cells and single-cell manipulation studies, although labor-intensive and technically challenging
	Virus-mediated delivery (AAV, Lentivirus)	Viral vectors deliver CRISPR/Cas9 genes into cells or target tissues for stable integration or expression	Suitable for long-term HBV gene editing studies, creation of stable HBV knockout cell lines
	Nanoparticles	Nanoparticles encapsulate CRISPR/Cas9 mRNAs, facilitating their uptake by cells	Efficient delivery of CRISPR/Cas9 for HBV gene editing in a wide range of cell types, suitable for high-throughput screening and functional studies
In vivo delivery	Virus-mediated delivery (AAV, Lentivirus)	Viral vectors are efficient carriers for delivering CRISPR/Cas9 components to target tissues	In vivo delivery of CRISPR/Cas9 for HBV gene editing in animal models, offering stable integration and long-term expression
	Nanoparticles	Encapsulate CRISPR/Cas9 components for efficient delivery to target tissues	In vivo delivery of CRISPR/Cas9 for HBV gene editing in liver tissues of animal models, with potential for clinical translation due to biocompatibility
	Hydrodynamic Injection	Rapid injection of a large volume of DNA solution induces transient transfection in the liver	Delivery of CRISPR/Cas9 for HBV gene editing in animal models, suitable for transient expression and rapid screening of gene editing efficacy
	in vivo electroporation	Brief electric pulses facilitate the uptake of CRISPR/Cas9 complexes by tissues	In vivo delivery of CRISPR/Cas9 for HBV gene editing in liver tissues of animal models, offering efficient transfection without viral vectors
	Microinjection	Direct injection of CRISPR/Cas9 components into target tissues or embryos	Precise delivery of CRISPR/Cas9 for HBV gene editing in animal models, suitable for studies requiring single-cell manipulation or spatial control