Protamine sulfate enhances lipid-mediated gene transfer
Protamine sulfate enhances lipid-mediated gene transfer
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DOI：
10.1038/sj.gt.3300484
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发表时间：
1997-09-01
期刊：
影响因子：
5.1
通讯作者：
Huang, L
中科院分区：
文献类型：
--
作者：
Sorgi, FL;Bhattacharya, S;Huang, L
聚阳离子肽，硫酸鱼精蛋白，USP，已经显示能够有效地浓缩质粒DNA，用于通过几种不同类型的阳离子脂质体在体外递送到几种不同类型的细胞中。单价阳离子脂质体制剂（DC-Chol和lipofectin）表现出与用多价阳离子脂质体制剂lipofectamine观察到的转染活性相当的增加的转染活性。这表明lipofectamine的上级体外活性源于其有效浓缩DNA的能力，并且鱼精蛋白的主要作用是浓缩剂，尽管它也具有类似于核定位信号的几个氨基酸序列。虽然先前已经报道了使用聚阳离子来缩合DNA，但是发现使用硫酸鱼精蛋白USP作为缩合剂上级聚-F-赖氨酸以及各种其他类型的鱼精蛋白。鱼精蛋白的各种盐形式之间的这些差异似乎可归因于鱼精蛋白之间的结构差异，而不是由于分子的净电荷的差异。鱼精蛋白分子内赖氨酸残基的出现与对质粒DNA的结合亲和力的降低以及观察到的转染增强活性的丧失相关。这一发现揭示了用于基因转移方案的缩合剂的结构要求。此外，硫酸鱼精蛋白，USP是FDA批准的具有记录的安全性特征的化合物，并且可以容易地用作人类基因治疗方案的佐剂。
A polycationic peptide, protamine sulfate, USP, has been shown to be able to condense plasmid DNA efficiently for delivery into several different types of cells in vitro by several different types of cationic liposomes. The monovalent cationic liposomal formulations (DC-Chol and lipofectin) exhibited increased transfection activities comparable to that seen with the multivalent cationic liposome formulation, lipofectamine. This suggests that lipofectamine's superior in vitro activity arises from ifs ability to condense DNA efficiently and that protamine's primary role is that of a condensation agent, although it also possesses several amino acid sequences resembling that of a nuclear localization signal. While the use of polycations to condense DNA has been previously reported, the use of protamine sulfate, USP as a condensation agent was found to be superior to poly-F-lysine as well as to various other types of protamine. These differences among various salt forms of protamine appear to be attributable to structural differences between the protamines and not due to differences in the net charge of the molecule. The appearance of lysine residues within the protamine molecule correlate with a reduction in binding affinity to plasmid DNA, as well as an observed loss in transfection-enhancing activity. This finding sheds light on the structural requirements of condensation agents for use in gene transfer protocols. Furthermore, protamine sulfate, USP is an FDA-approved compound with a documented safety profile and could be readily used as an adjuvant to a human gene therapy protocol.