组蛋白H2A显著增强体外DNA转染。
Histone H2A significantly enhances in vitro DNA transfection.
作者信息
Balicki D, Beutler E
机构信息
Scripps Research Institute, Department of Molecular and Experimental Medicine, La Jolla, California 92037, USA.
出版信息
Mol Med. 1997 Nov;3(11):782-7.
BACKGROUND
Gene transfer is a potential treatment modality of genetic disease. Efficient, practical methods of DNA transfection are currently under investigation.
MATERIALS AND METHODS
A beta-galactosidase reporter plasmid interacted electrostatically with histones, poly-L-Lys, poly-L-Arg, and a combination of poly-L-Lys and poly-L-Arg. This complex was then used to transfect COS-7 cells. beta-galactosidase activity was quantified and used to compare the efficiency of gene transfection in vitro. A comparison was also made of DNA transfection with the most active histone subclass, i.e., histone H2A, in the absence and presence of an anionic liposome.
RESULTS
There was a marked increase in DNA transfection in the presence of histone H2A when compared with the control, whereas each of the other histones and polycations showed little, if any, effect. The extent of activation depends strongly on the DNA/histone ratio and is also a function of the molarity of the final Tris-acetate, pH 8, solution. The anionic liposomes used demonstrated an inhibitory effect.
CONCLUSIONS
Histone H2A significantly enhances in vitro DNA transfection whereas other histones and anionic liposomes do not. A study of the difference between histone H2A and other histone subclasses may serve to clarify some of the mechanisms and the essential components of efficient gene delivery.
背景
基因转移是一种治疗遗传疾病的潜在方法。目前正在研究高效、实用的DNA转染方法。
材料与方法
β-半乳糖苷酶报告质粒与组蛋白、聚-L-赖氨酸、聚-L-精氨酸以及聚-L-赖氨酸和聚-L-精氨酸的组合发生静电相互作用。然后用该复合物转染COS-7细胞。对β-半乳糖苷酶活性进行定量，并用于比较体外基因转染的效率。还比较了在有无阴离子脂质体存在的情况下，用活性最高的组蛋白亚类即组蛋白H2A进行DNA转染的情况。
结果
与对照组相比，存在组蛋白H2A时DNA转染显著增加，而其他组蛋白和聚阳离子即使有影响也很小。激活程度强烈依赖于DNA/组蛋白比例，也是最终pH 8的Tris-乙酸盐溶液摩尔浓度的函数。所用的阴离子脂质体显示出抑制作用。
结论
组蛋白H2A显著增强体外DNA转染，而其他组蛋白和阴离子脂质体则不然。对组蛋白H2A与其他组蛋白亚类之间差异的研究可能有助于阐明高效基因传递的一些机制和基本成分。
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