I am interested in leaning about the structure prediction and design, molecular mechanisms and biological functions of siRNA, gene silencing (in special the PTK7 gene), and transfection using nanodevices (liposomes, polymer nanoparticules, etc.).
We have a few pages on the subject of siRNA transfection (e.g. https://altogen.com/transfection-resource/sirna-transfection/). For silencing experiments, you need to plan what protein you're targeting, what cell line you'll use, and the transfection reagent as well. Usually the transfection reagent is tailored to the cell line, and the cell line should preferably express high amounts of the protein of interest. As an example for PTK7, it has been silenced in HCT-116 in this study (Article Silencing of PTK7 in Colon Cancer Cells: Caspase-10-Dependen...
), and HCT-116 has its own designated transfection reagents as well (https://altogen.com/product/hct-116-transfection-reagent-colon-carcinoma/). There are a variety of online resources to also help you with siRNA design, and in particular the allele specificity necessary for successful silencing. You may want to look into siRNA library screening, as that can help you get a broader scope in your research.
Michael Davis What do you think about the
Calcium phosphate transfection for siRNA into lung cancer cells?
And about the PEG transfection (non-PEGylated) for siRNA into lung cancer cells? Are PEG-siRNA nanoparticles stable if PEG is not PEGylated with the siRNA molecule.
Andre Silva Pimentel, sorry for the delayed response. Calcium phosphate is an antiquated method of transfection, and is prone to cytotoxic effects and has generally lower efficiency than most other transfection techniques.
About PEG transfection, you'll still probably have stable siRNA nanoparticles with PEG, but those should be prepared right before the transfection. You should be fine if using PEG transfection reagents, as the lipid encapsulation process is generally unaffected by the presence of the PEG fragments.
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