Journal
/
Biology
/
siRNA Screening for å identifisere Ubiquitin og Ubiquitin-lignende system regulatorer av biologiske mekanismer i dyrkede pattedyrceller
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells

siRNA Screening for å identifisere Ubiquitin og Ubiquitin-lignende system regulatorer av biologiske mekanismer i dyrkede pattedyrceller

DOI:
10.3791/51572-v

•

10:43 min

•

May 24, 2014

•

, , , ,
1MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences,University of Dundee, 2Centre for Gene Regulation and Expression, College of Life Sciences,University of Dundee

Chapters

  • 00:05Title
  • 01:22Characterizing Hypoxia-responsiveness to Develop the Assay
  • 04:18Primary Screen: First Day
  • 07:27Primary Screen: Subsequent Days
  • 08:27Sample Screen
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

Her beskriver vi en metode for å utføre en målrettet siRNA "ubiquitome" skjermen for å identifisere romanen ubiquitin og ubiquitin-lignende regulatorer av HIF1A-mediert cellulær respons på hypoksi. Dette kan tilpasses til en hvilken som helst biologisk vei, hvor et robust lest ut av reporter-aktivitet er tilgjengelig.

Tags

SiRNA ScreeningUbiquitinUbiquitin-like SystemBiological PathwaysMammalian CellsPost-translational ModificationUbiquitomeHIF1AHypoxia ResponseTranscription-based Luciferase ReporterAssay DevelopmentPrimary ScreeningSecondary ScreeningTertiary/deconvolution ScreeningTargeted SiRNA LibraryOff-target Effects

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

MISSION esiRNA for RNAi Screening in Mammalian Cells
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

MISSION LentiPlex Samlede shRNA Library Screening i pattedyrceller
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

RNAi Screening for å identifisere Postembryonic fenotyper i C. elegans</em
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Isolering av ribosome Innbundne begynnende polypeptider<em> In vitro</em> For å identifisere translasjonelle Pause steder langs mRNA
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

En kvantitativ analyse for å studere Protein: DNA interaksjoner, Discover Transcriptional regulatorer Gene Expression, og identifisere nye anti-tumor Agenter
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Omvendt gjær to-hybrid system for å identifisere pattedyr Nuclear Receptor Rester som samhandler med Ligander og / eller antagonister
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Massivt Parallelle Reporter Analyser i dyrkede pattedyrceller
check_url/51572?article_type=v&slug=sirna-screening-to-identify-ubiquitin-ubiquitin-like-system

Imaging Lokal Ca<sup> 2+</sup> Signaler i dyrkede pattedyrceller

Read Article