I will be transfecting human and sheep endothelial cells with microRNA mimics and inhibitors. What is the best company to use for the mimics and inhibitors, and is there a particular vector I should be using? Also, is there a particular protocol which has worked for anyone?
May I ask which miR you're intending to mimic/inhibit? I did something similar a couple years ago in HeLa cells
If your approach is of using miRNA mimic/inhibitors integrated in viral mediated vector system, then I will wish u should get good answer from expert.
Otherwise, If you plan to transfect non-viral miRNA mimic/inhibitor to endothelial cells which are isolated from sheep and human, then there are pre-made mimics/inhibitors are available.
for mimic/inhibitor, I use Ambion (Thermo fisher) products. Still my preference will be 1. Ambion
2. Dharmacon
3. Eurogentec
4. Qiagen miR products.
While for transfection reagent, I use and recommend everyone the product from Polyplus (INTERferin), it works wonderfully since I struggled with transfection reagent for 3 month and why I am saying this because that particular branded product failed to transfect mimic/inhibitors to cancer cells, even many article also reported but don't know how they did.
yes, people uses Thermo RNAiMAX for transfecting.
For ref. see "Regulation of the Tissue Factor Isoform Expression and Thrombogenicity of HMEC-1 by miR-126 and miR-19a"
- DOI: 10.4172/2324-9293.1000101
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