Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems
Transfection efficiencies of several polymeric gene carriers were compared and correlated quantitatively to the amounts of cellular accumulation of plasmid DNA and to the expression of mRNA by quantitative real time PCR. Three cationic methacrylate polymer systems with similar chemical structure wer...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
2005
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| Subjects: | |
| Online Access: | http://hdl.handle.net/1721.1/30383 |
| _version_ | 1826194762206543872 |
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| author | Tan, J. F. Too, Heng-Phon Hatton, T. Alan Tam, K. C. |
| author_facet | Tan, J. F. Too, Heng-Phon Hatton, T. Alan Tam, K. C. |
| author_sort | Tan, J. F. |
| collection | MIT |
| description | Transfection efficiencies of several polymeric gene carriers were compared and correlated quantitatively to the amounts of cellular accumulation of plasmid DNA and to the expression of mRNA by quantitative real time PCR. Three cationic methacrylate polymer systems with similar chemical structure were used in this study, namely: poly(dimethylamino)ethyl methacrylate (PDMA) homopolymer, PEO-b-PDMA copolymer and PEO-b-poly(diethylamino)ethyl methacrylate (PEO-b-PDEA) copolymer. Despite their similar chemical structures, their transfection efficiencies were significantly different. PEO-b-PDEA copolymer was significantly less efficient as gene carrier compared to both PDMA and PEO-b-PDMA systems. Results from quantitative real-time polymerase chain reaction (real-time PCR), cytotoxicity and Zeta potential measurements showed correlations between the physical properties of the polymers and the efficiencies of cellular uptake of the transgene and transfections. In the case of PEO-b-PDEA system, cytotoxicity was due primarily to the excess polymers that did not participate in the DNA binding. In addition, the inability of the polymer/DNA complexes to interact with cell effectively was identified as the main barrier for high efficiency of transfection. This study demonstrated that the use of quantitative real-time PCR in combination with other physical characterization techniques can provide greater insights into the transfection barrier at different cellular levels. |
| first_indexed | 2024-09-23T10:01:44Z |
| format | Article |
| id | mit-1721.1/30383 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T10:01:44Z |
| publishDate | 2005 |
| record_format | dspace |
| spelling | mit-1721.1/303832019-04-12T08:15:24Z Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems Tan, J. F. Too, Heng-Phon Hatton, T. Alan Tam, K. C. Quantitative real time PCR transfection efficiency methacrylate polymer Transfection efficiencies of several polymeric gene carriers were compared and correlated quantitatively to the amounts of cellular accumulation of plasmid DNA and to the expression of mRNA by quantitative real time PCR. Three cationic methacrylate polymer systems with similar chemical structure were used in this study, namely: poly(dimethylamino)ethyl methacrylate (PDMA) homopolymer, PEO-b-PDMA copolymer and PEO-b-poly(diethylamino)ethyl methacrylate (PEO-b-PDEA) copolymer. Despite their similar chemical structures, their transfection efficiencies were significantly different. PEO-b-PDEA copolymer was significantly less efficient as gene carrier compared to both PDMA and PEO-b-PDMA systems. Results from quantitative real-time polymerase chain reaction (real-time PCR), cytotoxicity and Zeta potential measurements showed correlations between the physical properties of the polymers and the efficiencies of cellular uptake of the transgene and transfections. In the case of PEO-b-PDEA system, cytotoxicity was due primarily to the excess polymers that did not participate in the DNA binding. In addition, the inability of the polymer/DNA complexes to interact with cell effectively was identified as the main barrier for high efficiency of transfection. This study demonstrated that the use of quantitative real-time PCR in combination with other physical characterization techniques can provide greater insights into the transfection barrier at different cellular levels. Singapore-MIT Alliance (SMA) 2005-12-16T14:40:13Z 2005-12-16T14:40:13Z 2006-01 Article http://hdl.handle.net/1721.1/30383 en Molecular Engineering of Biological and Chemical Systems (MEBCS) 96110 bytes application/pdf application/pdf |
| spellingShingle | Quantitative real time PCR transfection efficiency methacrylate polymer Tan, J. F. Too, Heng-Phon Hatton, T. Alan Tam, K. C. Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title | Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title_full | Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title_fullStr | Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title_full_unstemmed | Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title_short | Correlating Gene Transfection Efficiency and the Physical Properties of Various Cationic Poly(methacrylate) Systems |
| title_sort | correlating gene transfection efficiency and the physical properties of various cationic poly methacrylate systems |
| topic | Quantitative real time PCR transfection efficiency methacrylate polymer |
| url | http://hdl.handle.net/1721.1/30383 |
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