کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2487553 | 1114422 | 2007 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Design of PCRâamplified DNA fragments for in vivo gene delivery: Sizeâdependency on stability and transgene expression
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موضوعات مرتبط
علوم پزشکی و سلامت
داروسازی، سم شناسی و علوم دارویی
اکتشاف دارویی
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چکیده انگلیسی
PCRâamplified DNA fragments can be more efficient and safer vectors than conventional plasmid DNA because of their smaller size and fewer numbers of immunostimulatory cytosineâphosphateâguanine (CpG) motifs. In the present study, the expression unit of plasmid DNA encoding farnesylated enhanced green fluorescent protein (EGFPF; pEGFPâF) or firefly luciferase (pLuc) was amplified by polymerase chain reaction (PCR) to obtain DNA fragments (EGFPFâmini, Lucâmini). EGFPFâmini was as effective as pEGFPâF on the basis of the number of EGFPFâexpressing cells after intravenous injection into mice by the hydrodynamicsâbased procedure. Then, the effects of the length of DNA fragments on transgene expression were examined using luciferaseâexpressing DNA preparations. Lucâmini preparations showed high levels of luciferase activity in cultured cells as well as in mouse liver, even although the levels did not exceed that of pLuc. An elongation of the DNA fragment on either side of the minimal expression unit was effective in increasing the transgene expression and the stability against nucleases. PCRâamplified DNA fragments showed a sustained luciferase activity in mouse liver compared with pLuc, indicating that they are effective in achieving a prolonged expression. Their stabilization against nucleases will further increase the potential of such short, structureâcontrolled and synthetic DNA fragments for in vivo gene delivery. © 2007 WileyâLiss, Inc. and the American Pharmacists Association J Pharm Sci 96: 2251-2261, 2007
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Pharmaceutical Sciences - Volume 96, Issue 9, September 2007, Pages 2251-2261
Journal: Journal of Pharmaceutical Sciences - Volume 96, Issue 9, September 2007, Pages 2251-2261
نویسندگان
Kazuhiro Hirata, Makiya Nishikawa, Naoki Kobayashi, Yuki Takahashi, Yoshinobu Takakura,