AI Article Synopsis
- We are creating effective techniques for gene transfer into tissue culture cells, using a combination of adenovirus and polylysine to enhance efficiency.
- We previously demonstrated that a chimeric adenovirus-polylysine-reporter-gene complex could successfully transfer a transporter gene into HeLa cells with high efficiency.
- Our latest methods involve simpler biochemical approaches that achieve nearly 100% transfection in various tissue culture cell lines, utilizing enzymatic coupling or biotin-streptavidin interactions to enhance the transfer process.
Article Abstract
We are developing efficient methods for gene transfer into tissue culture cells. We have previously shown that coupling of a chimeric adenovirus with polylysine allowed the construction of an adenovirus-polylysine-reporter-gene complex that transferred the transporter gene with great efficiency into HeLa cells. We have now explored simpler, biochemical means for coupling adenovirus to DNA/polylysine complexes and show that such complexes yield virtually 100% transfection in tissue culture cell lines. In these methods adenovirus is coupled to polylysine, either enzymatically through the action of transglutaminase or biochemically by biotinylating adenovirus and streptavidinylating the polylysine moiety. Combination complexes containing DNA, adenovirus-polylysine, and transferrin-polylysine have the capacity to transfer the reporter gene into adenovirus-receptor- and/or transferrin-receptor-rich cells.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC49445 | PMC |
| http://dx.doi.org/10.1073/pnas.89.13.6099 | DOI Listing |
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