[RNA Strand Displacement In Vitro]


Our team first decided to repeat the results of Winfree / Qian's 2011 paper on strand displacement, but using 2'-O-Methyl RNA tagged with fluorophores and quenchers. Figure A shows a foundational experiment performed on a platereader.
Figure A

Nucleic Acid Delivery


Delivery of Plasmid DNA to Mammalian Cells


Constitutive Expression of fluorescent markers etc
Delivery of 2'-O-Me RNA to Mammalian Cells

Movie of HEK293 cell being transfected with T*-S6*-ROX, as well as static images
15,20,25,30 pmol ratio data from FACS

Inducible Control of Protein Expression

DOX curve

Delivery of Plasmid DNA which transcribes short RNA Inputs

FF1 Knockdown Data

In Vivo RNA Strand Displacement

Strategy 1: Lipofectamine 2000 Transfection of RNA version of Reporter from Winfree/QIan 2011 Paper


Images will go here from April 24th experiment - display red fluorescence in all wells, including those that only got reporter or the wrong input - also see red vesicles indicating reporter comes apart inside the vesicles


Strategy 2: Switch Transfection reagent to RNAiMAX

RNAiMAX is supposed to be a better transfection reagent for double stranded RNA

Images will go here from experiment from June 13th onward where we do not see red vesicles, however we still see whole cell red fluorescence

Strategy 3: Tag RNA strand with an Alexa Fluorophore to act as a transfection marker

Strategy 4: Create DNA plasmids driving transcription of RNA inputs, while transfecting RNA Reporter

Strategy 4: Nucleofect RNA reporter, RNA inputs

[Strategy 5]: Redesign RNA Reporter