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Tips from the Bench: Transfection Tip

Deliver microRNA (miRNA) Effectively

Transfect miRNAs In Vitro Using the TransIT-TKO® and TransIT-siQUEST® Transfection Reagents

The TransIT-TKO and TransIT-siQUEST Transfection Reagents effectively transfect duplex miRNAs into mammalian cells in culture (Figure 1). These reagents, originally developed for siRNA transfection, efficiently transfect miRNAs due to their structural similarity to siRNAs. Knockdown efficiencies of these reagents may differ depending on the cell type. Ideally, both reagents should be tested to determine the best solution for your application. For miRNA transfection, follow each protocol and simply substitute duplex miRNA for siRNA.

A

Effectively Deliver miRNAs with TransIT TKO

B

Effectively Deliver miRNAs with TransIT siQUEST
Figure 1. Highly Effective Delivery of miRNA Duplexes Using TransIT-TKO® and TransIT-siQUEST® Transfection Reagents. Three miRNA reporter constructs were created in the psiCHECK™-2 Vector (Promega) by cloning the target sequences of miR-18, miR-143, and let7a within the 3’ untranslated region of Renilla luciferase mRNA. The psiCHECK™-2 vector also expresses the internal control, firefly luciferase. Each psiCHECK™ miRNA reporter plasmid was transfected into both HEK 293 and HeLa cells using TransIT®-LT1 Reagent. Approximately 6 hours post-transfection, the cells were transfected with 25 nM miR-18, miR-143 or let7a duplex miRNAs (Ambion) using either the (A) TransIT-TKO® or (B) TransIT-siQUEST® Transfection Reagents. Twenty-four hours post-miRNA transfection, luciferase expression was assayed. Renilla luciferase activity was normalized to firefly luciferase activity and compared to (-) control miRNA transfected samples.
 
 
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