Intracellular delivery can be achieved by bombarding cells or tissues with accelerated molecules or bacteria without the need for carrier particles.
To deliver non-permeable molecules into cells, one can utilize protocols such as microinjection, electroporation, liposome-mediated transfection or virus-mediated transfection. However, each method has its own limitations. Here we have developed a new molecular delivery technique where live cells or tissues are bombarded with highly accelerated molecules directly and without the need to conjugate the molecules onto carrier particles, which is essential in conventional "gene gun" experiments. Gene bombardments can be applied to well-differentiated cells, primary cultured cells/neurons or tissue explants, all of which are notoriously difficult to transfect. Exogenously made proteins and even bacteria can be effectively introduced into cells where they can execute their function or replicate. Our experimental results and physical model support the notion that accelerated chemicals, proteins, or microorganisms carry enough momentum to penetrate the plasma membrane. The bombardment process is associated with a transient (approximately 10 min) increase in cell permeability, but such membrane leakage has a minimal adverse effect on cell survival.
Lian
Wei-Nan
WN
Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.
Chang
Cheng-Hung
CH
Chen
Yin-Jhen
YJ
Dao
Ro-Lan
RL
Luo
Yun-Cin
YC
Chien
Jun-Yi
JY
Hsieh
Shie-Liang
SL
Lin
Chi-Hung
CH
D000818:Animals; D001419:Bacteria; D001431:Bacteriological Techniques; D019470:Biolistics / Q000379:methods*; D016466:CHO Cells; D002460:Cell Line; D002463:Cell Membrane Permeability; D006224:Cricetinae; D003412:Cricetulus; D003598:Cytoskeletal Proteins / Q000008:administration & dosage / Q000737:chemistry; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000235:genetics; D016503:Drug Delivery Systems / Q000379:methods*; D004926:Escherichia coli; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000737:chemistry; D014162:Transfection
D003598:Cytoskeletal Proteins; D004274:DNA, Recombinant; D011994:Recombinant Proteins