Katakura Hiromichi, Harada Atsushi, Kataoka Kazunori, Furusho Miki, Tanaka Fumihiro, Wada Hiromi, Ikenaka Kazuhiro
National Institute for Physiological Sciences, Okazaki National Research Institutes, Okazaki, Japan.
J Gene Med. 2004 Apr;6(4):471-7. doi: 10.1002/jgm.519.
Although some cationic reagents, such as polybrene, improve gene transduction in vitro, their use in vivo is prohibited due to their toxicity to the exposed cells. This paper demonstrates that a new cationic reagent, poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLL), improves gene transduction with retroviral vectors without increasing cell toxicity.
A retroviral vector derived from the Moloney leukemia virus, containing the lacZ gene, was modified with PEG-PLL prior to transduction into NIH3T3, Lewis lung carcinoma, and primary cultured mouse brain cells. LacZ transduction efficacy was evaluated by counting the number of X-Gal-positive cells.
We have demonstrated that PEG-PLL is able to stably modify the viral particle surface due to the affinity of the PEG moiety to the biomembrane, and neutralizes negative charges by the cationic nature of the poly-lysine residue. Thus, PEG-PLL increased the gene transduction efficiency and minimized cell toxicity because free PEG-PLL was removable by centrifugation. We have shown that PEG-PLL increased the viral gene transduction efficiency 3- to 7-fold with NIH3T3 or Lewis lung carcinoma cell lines without increasing cytotoxicity. It improved retroviral gene transduction efficacy even against labile cells, such as primary cultured brain cells.
PEG-PLL is a novel reagent that improves retroviral gene transduction efficacy without increasing cytotoxicity.
尽管一些阳离子试剂，如聚凝胺，可在体外提高基因转导效率，但因其对暴露细胞的毒性，禁止在体内使用。本文证明，一种新型阳离子试剂，聚（乙二醇）-聚（L-赖氨酸）嵌段共聚物（PEG-PLL），可在不增加细胞毒性的情况下，提高逆转录病毒载体的基因转导效率。
将来源于莫洛尼白血病病毒、含有lacZ基因的逆转录病毒载体，在转导至NIH3T3细胞、Lewis肺癌细胞和原代培养的小鼠脑细胞之前，用PEG-PLL进行修饰。通过计数X-Gal阳性细胞的数量来评估LacZ转导效率。
我们已经证明，由于PEG部分对生物膜的亲和力，PEG-PLL能够稳定地修饰病毒颗粒表面，并通过聚赖氨酸残基的阳离子性质中和负电荷。因此，PEG-PLL提高了基因转导效率，并将细胞毒性降至最低，因为游离的PEG-PLL可通过离心去除。我们已经表明，PEG-PLL可使NIH3T3或Lewis肺癌细胞系的病毒基因转导效率提高3至7倍，而不增加细胞毒性。它甚至对不稳定细胞，如原代培养的脑细胞，也能提高逆转录病毒基因转导效率。
PEG-PLL是一种新型试剂，可在不增加细胞毒性的情况下提高逆转录病毒基因转导效率。
