Gap junction mediated transport of shRNA between human embryonic stem cells.
Gap junction mediated transport of shRNA between human embryonic stem cells.
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DOI：
10.1016/j.bbrc.2007.09.035
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发表时间：
2007-11
影响因子：
3.1
通讯作者：
E. Wolvetang;M. Pera;K. Zuckerman
中科院分区：
文献类型：
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作者：
E. Wolvetang;M. Pera;K. Zuckerman
间隙连接细胞内通讯（GJIC）允许小分子在相邻细胞之间直接转运。我们假设一个hESC中的siRNAs可以通过GJIC抑制另一个hESC中靶RNA的表达。我们将表达绿色荧光蛋白（GFP）的ENVY hESC与不表达GFP的hESC共培养，后者已被转导以稳定表达针对GFP的shRNA。我们发现GFP shRNA表达的hESC以剂量依赖的方式抑制相邻GFP表达的hESC中GFP的表达。当用两个核苷酸突变的非功能性GFP shRNA转导共培养的细胞时，或者当细胞与GJIC抑制剂α-甘草次酸（α-GA）孵育时，ENVY细胞中GFP表达的下调未观察到。我们的结论是，21- 23 bp的双链shRNA/siRNA寡核苷酸能够移动通过hESC之间的间隙连接，从而可以影响相邻hESC中的基因表达。这种新的细胞间基因表达调控机制可能为hESC的调控提供新的途径。
Gap junction intracellular communication (GJIC) allows the direct transport of small molecules between adjacent cells. We hypothesized that siRNAs in one hESC could inhibit target RNA expression in another hESC via GJIC. We co-cultured green fluorescent protein (GFP)-expressing ENVY hESC with non-GFP-expressing hESC, which had been transduced to stably express shRNA directed against GFP. We discovered that the GFP shRNA expressing hESC inhibited GFP expression in the adjacent GFP-expressing hESC in a dose-dependent manner. This downregulation of GFP expression in ENVY cells was not observed when the co-cultured cells had been transduced with a non-functional GFP shRNA that was mutated at two nucleotides or when the cells were incubated with the GJIC inhibitor, α-glycyrrhetinic acid (α-GA). We conclude that 21–23bp double-stranded shRNA/siRNA oligonucleotides are able to move through gap junctions between hESCs and thus can affect gene expression in neighbouring hESC. This novel intercellular gene expression regulatory mechanism may offer new approaches to manipulation of hESC.