Jul 07, 2023

Public workspacesiRNA-mediated knockdown of TFE3

DOI
dx.doi.org/10.17504/protocols.io.3byl4q26rvo5/v1
  • Narayana Yadavalli1,2,3,4,5,
  • Shawn M. Ferguson1,2,3,4,5,6
  • 1Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA;
  • 2Department of Neuroscience, Yale University School of Medicine, New Haven, Connecticut 06510, USA;
  • 3Program in Cellular Neuroscience, Neurodegeneration and Repair;
  • 4Wu Tsai Institute Yale University School of Medicine, New Haven, Connecticut 06510, USA;
  • 5Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815, USA;
  • 6Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, Connecticut 06510, USA
Berrak Ugur
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DOI: dx.doi.org/10.17504/protocols.io.3byl4q26rvo5/v1
Protocol Citation: Narayana Yadavalli, Shawn M. Ferguson 2023. siRNA-mediated knockdown of TFE3. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl4q26rvo5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 23, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 83917
Keywords: siRNA-mediated knockdown of TFE3, ASAPCRN
Funders Acknowledgement:
ASAP
Grant ID: 000580
Abstract
This protocol details siRNA-mediated knockdown of TFE3.
Attachments
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Materials
Reagents required

  • RNAiMAX (Invitrogen)
  • Opti-MEM (Invitrogen)
siRNA-mediated knockdown of TFE3
siRNA-mediated knockdown of TFE3
1h 30m
1h 30m
On the day before transfection, plate 100,000 cells per well in a 6 well dish.
Change media Duration01:00:00 before transfection (Amount2 mL with no antibiotics).
1h
The next day, assemble the siRNA transfection reaction by combining Amount7.5 µL of Concentration20 micromolar (µM) siRNA, Amount5 µL RNAiMAX transfection reagent (Invitrogen), and Amount500 µL Opti-MEM (Invitrogen) in a sterile 1.7ml tube.
Pipetting
Incubate transfection mix at TemperatureRoom temperature for Duration00:30:00 .

30m
Incubation
After incubation, add the transfection mixture drop wise to the cells.
Pipetting
Collect the lysates for immunoblotting 2 days post-transfection.