Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells.
Synthetic vectors were evaluated for their ability to mediate efficient mRNA transfection. Initial results indicated that lipoplexes, but not polyplexes based on polyethylenimine (PEI, 25 and 22 kDa), poly(L-lysine) (PLL, 54 kDa) or dendrimers, mediated efficient translation of mRNA in B16-F10 cells...
| প্রধান লেখক: | , , , , |
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| বিন্যাস: | Journal article |
| ভাষা: | English |
| প্রকাশিত: |
2001
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| _version_ | 1826291102132469760 |
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| author | Bettinger, T Carlisle, R Read, M Ogris, M Seymour, L |
| author_facet | Bettinger, T Carlisle, R Read, M Ogris, M Seymour, L |
| author_sort | Bettinger, T |
| collection | OXFORD |
| description | Synthetic vectors were evaluated for their ability to mediate efficient mRNA transfection. Initial results indicated that lipoplexes, but not polyplexes based on polyethylenimine (PEI, 25 and 22 kDa), poly(L-lysine) (PLL, 54 kDa) or dendrimers, mediated efficient translation of mRNA in B16-F10 cells. Significant mRNA transfection was achieved by lipoplex delivery in quiescent (passage 0) human umbilical vein endothelial cells (HUVEC), and by passage 4, 10.7% of HUVEC were transfected compared to 0.84% with DNA. Lack of expression with PEI 25 kDa/mRNA or PLL 54 kDa/mRNA in a cell-free translation assay and following cytoplasmic injection into Rat1 cells indicated that these polyplexes were too stable to release mRNA. In contrast, polyplexes formed using smaller PEI 2 kDa and PLL 3.4 kDa gave 5-fold greater expression in B16-F10 cells compared to DOTAP, but were dependent on chloroquine for transfection activity. Endosomolytic activity was incorporated by conjugating PEI 2 kDa to melittin and resulting PEI 2 kDa-melittin/mRNA polyplexes mediated high transfection levels in HeLa cells (31.1 +/- 4.1%) and HUVEC (58.5 +/- 2.9%) in the absence of chloroquine, that was potentiated to 52.2 +/- 2.7 and 71.6 +/- 1.7%, respectively, in the presence of chloroquine. These results demonstrate that mRNA polyplexes based on peptide-modified low molecular weight polycations can possess versatile properties including endosomolysis that should enable efficient non-viral mRNA transfection of quiescent and post-mitotic cells. |
| first_indexed | 2024-03-07T02:54:21Z |
| format | Journal article |
| id | oxford-uuid:aec4706c-0281-4c4f-9b19-647e3290197c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:54:21Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:aec4706c-0281-4c4f-9b19-647e3290197c2022-03-27T03:44:57ZPeptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:aec4706c-0281-4c4f-9b19-647e3290197cEnglishSymplectic Elements at Oxford2001Bettinger, TCarlisle, RRead, MOgris, MSeymour, LSynthetic vectors were evaluated for their ability to mediate efficient mRNA transfection. Initial results indicated that lipoplexes, but not polyplexes based on polyethylenimine (PEI, 25 and 22 kDa), poly(L-lysine) (PLL, 54 kDa) or dendrimers, mediated efficient translation of mRNA in B16-F10 cells. Significant mRNA transfection was achieved by lipoplex delivery in quiescent (passage 0) human umbilical vein endothelial cells (HUVEC), and by passage 4, 10.7% of HUVEC were transfected compared to 0.84% with DNA. Lack of expression with PEI 25 kDa/mRNA or PLL 54 kDa/mRNA in a cell-free translation assay and following cytoplasmic injection into Rat1 cells indicated that these polyplexes were too stable to release mRNA. In contrast, polyplexes formed using smaller PEI 2 kDa and PLL 3.4 kDa gave 5-fold greater expression in B16-F10 cells compared to DOTAP, but were dependent on chloroquine for transfection activity. Endosomolytic activity was incorporated by conjugating PEI 2 kDa to melittin and resulting PEI 2 kDa-melittin/mRNA polyplexes mediated high transfection levels in HeLa cells (31.1 +/- 4.1%) and HUVEC (58.5 +/- 2.9%) in the absence of chloroquine, that was potentiated to 52.2 +/- 2.7 and 71.6 +/- 1.7%, respectively, in the presence of chloroquine. These results demonstrate that mRNA polyplexes based on peptide-modified low molecular weight polycations can possess versatile properties including endosomolysis that should enable efficient non-viral mRNA transfection of quiescent and post-mitotic cells. |
| spellingShingle | Bettinger, T Carlisle, R Read, M Ogris, M Seymour, L Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title | Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title_full | Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title_fullStr | Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title_full_unstemmed | Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title_short | Peptide-mediated RNA delivery: a novel approach for enhanced transfection of primary and post-mitotic cells. |
| title_sort | peptide mediated rna delivery a novel approach for enhanced transfection of primary and post mitotic cells |
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