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1. What is shRNA?
shRNA stands for short-haiprin RNA which is an artificial RNA molecule with a tight hairpin turn that can be used to silence target gene expression via RNA interference (RNAi). shRNA is an advantageous mediator of RNAi in that it has a relatively low rate of degradation and turnover.
2. What are the advantages of using shRNAs versus siRNAs?
siRNAs are chemically synthesized and their use is restricted to experiments studying the impact of transient suppression of gene expression. shRNAs are carried within the context of a plasmid or viral-based vector. The vector may carry an antibiotic-resistance gene or a reporter gene, which permits the selection of a stably transfected cell population.
3. What is the difference between using shRNA viruses versus shRNA plasmids?
Both shRNA plasmids and shRNA viruses may be used to develop stable expression of the shRNA with antibiotic selection. Using viral-based shRNA delivery vectors can efficiently deliver the shRNA into virtually any mammalian cell type including that are difficult (or impossible) to transfect. Additionally, viral transduction is a much more efficient process than transfection.
4. What is the function of the scrambled control?
The scrambled control clone is constructed by cloning a scrambled sequence (one that does not match any genomic sequences) into shRNA vectors. It serves as a negative control to eliminate the potential non-specific effect induced by expression of the plasmid.
5. What is Creative Biogene’s guarantee for the shRNA clones?
Creative Biogene guarantees that the shRNA sequences in the expression cassettes are identical to the target gene. In most cases, we can guarantee at least one construct produce a 70% or greater knockdown efficiency determined by qRT-PCR.
6. What type of promoter does your vector use for shRNA transcription?
The vector uses promoter H1, or U6, or EF-1a etc.