A simple method for delivering morpholino antisense oligos into the cytoplasm of cells
A simple method for delivering morpholino antisense oligos into the cytoplasm of cells
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DOI：
10.1089/oli.1.1996.6.169
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发表时间：
1996-09-01
期刊：
影响因子：
--
通讯作者：
Summerton, J
中科院分区：
文献类型：
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作者：
Partridge, M;Vincent, A;Summerton, J
我们报道了一种简单而有效的方法，用于将吗啉代反义寡核苷酸递送到培养的贴壁依赖性动物细胞的胞质溶胶中。这种方法，称为刮片加载，在几秒钟内进行，使用普通廉价的实验室工具，并且对细胞的有害影响最小。使用这种递送方法，在细胞外培养基中以0.1 μ M和1 μ M存在的吗啉代寡核苷酸分别以56%和85%的水平抑制其在培养的Hela细胞内的靶向遗传序列。两种浓度高达3 μ M的对照吗啉代寡核苷酸没有抑制作用，表明这种结构类型具有良好的序列特异性。还描述了一种用于培养细胞中反义活性的简单、快速和灵敏定量的测试系统。
We report a simple and effective means for delivering Morpholino antisense oligos into the cytosol of cultured anchorage-dependent animal cells, This method, referred to as scrape-loading, is carried out in a matter of seconds, uses a common inexpensive laboratory implement, and has minimal detrimental impact on the cells, Using this delivery method, a Morpholino oligo present at 0.1 mu M and 1 mu M in the extracellular medium inhibited its targeted genetic sequence within cultured Hela cells at levels of 56% and 85%, respectively. Lack of inhibition by two control Morpholino oligos at concentrations up to 3 mu M indicates good sequence specificity by this structural type, Also described is a test system for simple, rapid, and sensitive quantitation of antisense activity in cultured cells.