shRNA-Induced Gene Knockdown In Vivo to Investigate Neutrophil Function.
shRNA-Induced Gene Knockdown In Vivo to Investigate Neutrophil Function.
复制标题
shRNA 诱导体内基因敲低以研究中性粒细胞功能。
DOI：
10.1007/978-1-4939-3480-5_13
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Wu,Dianqing
中科院分区：
文献类型：
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作者：
Basit,Abdul;Tang,Wenwen;Wu,Dianqing
为了有效地沉默中性粒细胞中的基因，我们利用RNA干扰并开发了基于shRNA的基因敲除技术。该方法涉及用携带针对特定基因的shRNA的逆转录病毒载体转染小鼠骨髓来源的造血干细胞。然后将转染的干细胞移植到受辐射的野生型小鼠中。在干细胞植入后，移植的小鼠具有两组循环中性粒细胞。一组具有被敲除的感兴趣的基因，而另一组具有表达基因的完全互补。这种有效的技术提供了一种独特的方法，直接比较在相同环境中具有敲低基因的中性粒细胞与具有完整表达基因的中性粒细胞的反应。
To silence genes in neutrophils efficiently, we exploited the RNA interference and developed an shRNA-based gene knockdown technique. This method involves transfection of mouse bone marrow-derived hematopoietic stem cells with retroviral vector carrying shRNA directed at a specific gene. Transfected stem cells are then transplanted into irradiated wild-type mice. After engraftment of stem cells, the transplanted mice have two sets of circulating neutrophils. One set has a gene of interest knocked down while the other set has full complement of expressed genes. This efficient technique provides a unique way to directly compare the response of neutrophils with a knocked-down gene to that of neutrophils with the full complement of expressed genes in the same environment.