{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T06:27:42Z","timestamp":1790922462143,"version":"4.1.0"},"reference-count":52,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2003,4,30]],"date-time":"2003-04-30T00:00:00Z","timestamp":1051660800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2003,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Elevated plasma high\u2010density lipoprotein (HDL), and its major constituent apolipoprotein AI (apoAI), are cardioprotective. Paradoxically, two natural variants of apoAI, termed apoAI<jats:sub>Milano<\/jats:sub> and apoAI<jats:sub>Paris<\/jats:sub>, are associated with low HDL, but nevertheless provide remarkable protection against heart disease for heterozygous carriers and may even lead to longevity. Both variants arise from point mutations and have Arg<jats:sup>173<\/jats:sup> and Arg<jats:sup>151<\/jats:sup> to Cys substitutions, respectively, which allow disulphide\u2010linked dimers to form. Potentially, synthetic RNA\/DNA oligonucleotides (chimeraplasts) can permanently correct single point mutations in genomic DNA. Here, we use a variation of such targeted gene repair technology, \u2018gain\u2010of\u2010function chimeraplasty\u2019, and attempt to enhance the biological activity of apoAI by altering a single genomic base to generate the atheroprotective phenotypes, apoAI<jats:sub>Milano<\/jats:sub> and apoAI<jats:sub>Paris<\/jats:sub>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We targeted two cultured cell lines that secrete human apoAI, hepatoblastoma HepG2 cells and recombinant CHO\u2010AI cells, using standard 68\u2010mer chimeraplasts with polyethyleneimine (PEI) as carrier and then systematically varied several experimental conditions. As a positive control we targeted the dysfunctional <jats:italic>APOE2<\/jats:italic> gene, which we have previously converted to wild\u2010type <jats:italic>APOE3<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Conversion of wild\u2010type apoAI to apoAI<jats:sub>Milano<\/jats:sub> proved refractory, with limited correction in CHO\u2010AI cells only. However, a successful conversion to apoAI<jats:sub>Paris<\/jats:sub> was achieved, as demonstrated by polymerase chain reaction\u2010restriction fragment length polymorphism (PCR\u2010RFLP) analysis and direct genomic sequencing. Unexpectedly, attempts with a new batch of 68\u2010mer chimeraplast to enhance conversion, by using different delivery vehicles, including chemically modified PEI, failed to show a base change; nor could conversion be detected with an 80\u2010mer or a 52\u201376\u2010mer series. In contrast, when a co\u2010culture of CHO\u2010E2 and CHO\u2010AI cells was co\u2010targeted, a clear conversion of apoE2 to apoE3 was seen, whereas no apoAI<jats:sub>Paris<\/jats:sub> could be detected. When the individual chimeraplasts were analysed by denaturing electrophoresis only the active apoE2\u2010to\u2010E3 chimeraplast gave a sharp band.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our findings suggest that different batches of chimeraplasts have variable characteristics and that their quality may be a key factor for efficient targeting and\/or base conversion. We conclude that, although an evolving technology with enormous potential, chimeraplast\u2010directed gene repair remains problematical. Copyright \u00a9 2003 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.403","type":"journal-article","created":{"date-parts":[[2003,8,27]],"date-time":"2003-08-27T13:51:03Z","timestamp":1061992263000},"page":"795-802","source":"Crossref","is-referenced-by-count":14,"title":["Failure to generate atheroprotective apolipoprotein AI phenotypes using synthetic RNA\/DNA oligonucleotides 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