Intracellular delivery can be achieved by bombarding cells or tissues with accelerated molecules or bacteria without the need for carrier particles |
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Authors: | Lian Wei-Nan Chang Cheng-Hung Chen Yin-Jhen Dao Ro-Lan Luo Yun-Cin Chien Jun-Yi Hsieh Shie-Liang Lin Chi-Hung |
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Institution: | Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan. |
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Abstract: | 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. |
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Keywords: | F-actin filamentous actin IF immunofluorescence D3K dextran (MW:3K) D40K dextran (MW:40K) D70K dextran (MW:70K) D500K dextran (MW:500K) CHO cell Chinese Hamster Ovary cell |
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