Journal
/
Genetics
/
工程人为因素来特异性地操纵人细胞中的选择性剪接
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
Please note that all translations are automatically generated. Click here for the English version.

工程人为因素来特异性地操纵人细胞中的选择性剪接

DOI:
10.3791/54967-v

•

10:06 min

•

April 26, 2017

•

, , , , , ,
1Key Laboratory of Computational Biology, CAS-MPG Partner Institute for Computational Biology,Shanghai Institutes for Biological Sciences (SIBS), 2Institute of Cancer Stem Cell, Second Affiliated Hospital, Cancer Center,Dalian Medical University

Chapters

  • 00:05Title
  • 00:51Construction of the PUF RNA-binding Domain Scaffold with Customized RNA Binding Specificity by Overlapping PCR
  • 02:05Transfection of Engineered Splicing Factors (ESFs) for Modulation of Exon Inclusion and Alternative Splice-site Use
  • 03:43Cell Harvest and Identification of Spliced Isoforms
  • 06:24Modulation of Endogenous Bcl-x Splicing and Apoptosis by ESFs
  • 08:23Results: Subcellular Localization of ESFs in Transfected HeLa Cells
  • 09:30Conclusion

Summary

Automatic Translation

Automatic Translation

该报告介绍了一种生物工程方法来设计和构建特异性调节在哺乳动物细胞中靶基因的剪接新颖人工剪接因子（ASFS）。该方法可以进一步扩展来设计各种人为因素来操纵RNA代谢的其它方面。

Tags

Alternative SplicingArtificial Splicing FactorsPUF DomainRNA ProcessingHEK-293T CellsTransfectionSplicing Reporter PlasmidsGlycine-rich DomainRS-effector DomainPGL-RS-PUF Expression Vector

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

在新鲜分离的人代绒毛滋养细胞转染和EMSA分析

暴露在生理条件下的氧气在人体细胞中帽结合蛋白质分析

CRISPR/Cas9 对小鼠和人造血祖细胞的高效基因破坏

RNA原位杂交法定量分析小鼠脑切片中的替代前 mRNA 拼接方法

CRISPR/Cas9 诱导人多潜能干细胞内源蛋白标记的应用

基于细胞的 sineup 非编码 rna 检测, 可专门增强 mrna 的翻译能力

基于细菌选择和生长的盘状细胞基因工程

用于分离共转录RNA处理事件的人工RNA聚合酶II伸长复合物

伦蒂病毒介导基因在低附着板中制备的人类伪性沉默

哺乳动物细胞中基于CRISPR的聚集基因屏幕

Read Article