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在哺乳动物细胞的RNAi筛选使命esiRNA
JoVE Journal
Biology
This content is Free Access.
JoVE Journal
Biology
MISSION esiRNA for RNAi Screening in Mammalian Cells
Please note that all translations are automatically generated.
Click here for the English version.
在哺乳动物细胞的RNAi筛选使命esiRNA
DOI:
10.3791/2008-v
•
15:31 min
•
May 12, 2010
•
Mirko Theis
,
Frank Buchholz
1
Max Planck Institute of Molecular Cell Biology and Genetics
Chapters
00:00
Title
01:43
Introduction
02:22
Use of High-Quality MISSION esiRNA Libraries for RNAi Screening
03:49
Optimizing Transfection Prior to Screening
05:28
Setting Up the Assay and Preparing for the Primary Screen
09:34
The Primary Screen
12:03
Secondary Screen of the Selected Genes for Hit Validation
13:40
Representative Results Screening for Mammalian Cell Cycle Genes Using MISSION esiRNA
14:43
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
这里我们使用一个参与细胞分裂的基因的高通量筛选人类esiRNA库。我们演示了如何设置和行为的一个esiRNA屏幕,以及如何分析和验证的结果。
Tags
MISSION EsiRNA
RNAi Screening
Mammalian Cells
Gene Expression Control
Small Interfering RNAs (siRNAs)
Dicer
RNA-induced Silencing Complex (RISC)
Target MRNA Cleavage
Loss Of Gene Function Studies
Chemical Synthesis Of SiRNAs
Endonucleolytic Cleavage
Target-specific Long Double-stranded RNAs
RNase III
Endoribonuclease-prepared SiRNA
Efficacy Of SiRNAs And EsiRNAs
Off-target Effects
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