RNA interference (RNAi) is an experimental technique in which a small (19 -23bp) exogenous double-stranded RNA (siRNA) homologous to the coding region of an endogenous mRNA is introduced into a cell, triggering efficient and specific degradation of the corresponding mRNA in the cell, resulting in the silencing of specific gene expression (knock down). BOC Sciences offers a service to test the effects of RNAi interference. We use real-time quantitative PCR to detect knock down effects at the mRNA level and Western blot to detect knock down effects at the protein level.
Gene silencing is an important form of gene expression regulation. It is mainly manifested as down-regulation of the expression of a specific gene or inhibition of expression. There are two main aspects of gene silencing: gene silencing at the transcriptional level and gene silencing at the post-transcriptional level. The former is mainly caused by gene methylation, heterochromatinization and positional effects. The latter is caused by antisense RNA and RNA interference. Among them, RNA interference technology has shown great application prospects in gene function research and drug development in the post-genome era by efficient and specific target gene inhibition.
It is a phenomenon of silencing of homologous gene expression induced by double-stranded RNA. In RNA interference experiments, the selection of efficient siRNAs is critical for successful silencing or shutting down of gene expression as the efficiency of gene silencing mediated by siRNAs is unpredictable. Both the detection of the silencing effect of specific siRNAs and the selection of efficient siRNAs can be verified by bio-experimental means of real-time quantitative PCR or Western blot.
Factors influencing the efficiency of siRNA interference include:
The experimental report includes a siRNA sequence, a sequencing report of constructed vector, a fluorescent quantitative PCR detection report, Western Blot pictures, all experimental data and detailed experimental steps.
siRNA interference detection evaluates how effectively a specific siRNA or shRNA silences the target gene at the mRNA and protein levels, ensuring reliable experimental results.
Real-time quantitative PCR is used to assess mRNA knockdown, while Western blot detects reduction at the protein level.
Typically, 3 siRNA target sequences (19-21 nt each) are designed per gene to maximize the likelihood of achieving efficient gene silencing.
Efficiency can be influenced by shRNA vector design, target gene properties (secondary structure, microenvironment), cell type, transfection efficiency, transfection reagents, and protein half-life.
