Introduction of liposome-encapsulated SV40 DNA into cells.
Introduction of liposome-encapsulated SV40 DNA into cells.
复制标题
DOI：
10.1016/s0021-9258(19)70482-7
复制
发表时间：
1980-11
期刊：
影响因子：
--
通讯作者：
R. Fraley;S. Subramani;P. Berg;D. Papahadjopoulos
中科院分区：
文献类型：
--
作者：
R. Fraley;S. Subramani;P. Berg;D. Papahadjopoulos
从猿猴病毒40（SV 40）中分离的DNA被包裹在大的（0.4微米直径）单层磷脂囊泡中。用于脂质体制备的程序以高效率（30至50%包封）包封SV 40 DNA，并且不改变DNA分子的物理或生物学性质。脂质体包埋的病毒DNA的生物活性通过在允许的猴细胞系上的噬斑测定来确定。脂质体包埋的SV 40 DNA的感染性比游离裸DNA的感染性增强至少100倍。重要的是，囊泡包埋的DNA的感染性是耐DNA酶消化，依赖于每个囊泡和囊泡脂质组合物上封装的DNA的量。由磷脂酰丝氨酸组成的脂质体是最有效的DNA递送细胞（1.8 × 10（3）空斑形成单位/微克DNA）。将含有DNA的脂质体与细胞一起孵育后，通过将细胞暴露于高浓度的聚乙二醇或甘油，其感染性可额外增强10至200倍。在这些条件下，脂质体包封的SV 40 DNA（3 × 10（5）空斑形成单位/微克）的感染性与使用磷酸钙法报告的值相当。除了提供用于监测和优化囊泡内容物向细胞的递送的灵敏测定外，脂质体介导的核酸递送可能具有增加DNA向细胞递送的效率和扩大可被转化或转染的细胞类型的数量的潜力。
DNA, isolated from Simian virus 40 (SV40), has been encapsulated in large (0.4-micrometer diameter) unilamellar phospholipid vesicles. The procedure used for liposome preparation encapsulated the SV40 DNA at high efficiency (30 to 50% entrapment) and did not alter the physical or biological properties of the DNA molecules. The biological activity of the liposome-entrapped viral DNA was determined by plaque assays on a permissive monkey cell line. The infectivity of liposome-entrapped SV40 DNA was enhanced at least 100-fold over that of free naked DNA. Importantly, the infectivity of vesicle-entrapped DNA was resistant to DNase digestion, dependent on the amount of DNA encapsulated per vesicle and on the vesicle lipid composition. Liposomes composed of phosphatidylserine were the most efficient for delivery of DNA to cells (1.8 x 10(3) plaque-forming units/micrograms of DNA). Following the incubation of DNA-containing liposomes with cells, their infectivity could be enhanced an additional 10- to 200-fold by exposing the cells to high concentrations of polyethylene glycol or glycerol. Under these conditions the infectivity of liposome-encapsulated SV40 DNA (3 x 10(5) plaque-forming units/microgram) was comparable with values reported using the calcium phosphate method. In addition to providing a sensitive assay for monitoring and optimizing the delivery of vesicle contents to cells, the liposome-mediated delivery of nucleic acids may have potential for increasing the efficiency of DNA delivery to cells and for extending the number of cell types which can be transformed or transfected.