Can someone give the brief of siRNA knock down procedure of a gene? How does it work? I want to check the role of one of the protein in some of the breast cancer cell line. For this ist i am gonna use SiRNA knock down of gene for that protein, in order to check whether knock down/interference of this gene can inhibit the cancer proliferation or not? Is this the right approach? Can anyone give me the brief detail of the procedure of siRNA interference? Thanks in advance.
Here you'll find an interesting webinar about siRNAs.
https://app1.horizondiscovery.com/webinar-getting-started-sirna-considerations-recommendations-register
We usually use ON-TARGETplus siRNA smart pools from Dharmacon/Horizon (a pool of different siRNA sequences targeting the same gene) for achieving a better "knock-down" efficiency (assessed by qPCR). It is definitely a a method you can use for your assay to asses if cell proliferation is altered upon silencing your target gene. Nevertheless, we usually use additional methods such as stable shRNA mediated knockout and or /CRISPR/Cas9 mediated ko to verify the results obtained by siRNA interference with additional methods.
I advise you to order predesigned validated siRNA. The optimal solution is to use pooled siRNA - a mix of 3-4 sequences. Using a mix enchances efficiency of a knock down and reduces off target effect because concentration of individual sequences is lower than when you use single siRNA.
Sascha Seidel Magdalena Oroń Thank you so much. Your answers are helpful in clearing my confusion.
Article Functional siRNAs and miRNAs exhibit strand bias (vol 115, p...
Hi Dear
Please read this article.
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