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Gene knockdown cell lines play important roles for studying gene function, designing diseases models, biopharmaceutical research, drug discovery and many other applications. shRNA is an advantageous mediator of RNAi in that it has a relatively low rate of degradation and turnover. Scientists of Creative Biogene have developed a series of gene knockdown stable cell lines to assist your research.
Advantages
RNA interference (RNAi) refers to a phenomenon that is highly conserved during evolution, induced by double-stranded RNA, and causes efficient and specific degradation of homologous mRNA. At the cellular level, RNAi technology is used to inhibit the expression of target genes to obtain gene knockdown cell lines, which have been widely used in studying gene functions, designing disease models, and drug development.
Applications for gene knockdown stable cell lines include:
(1) Studying gene functions: By specifically targeting and knocking down a specific gene, researchers can observe the effects of the gene's absence on cellular processes. This provides valuable insights into the role of this gene in various biological pathways. For example, knockdown genes involved in cell cycle regulation can help determine their importance in controlling cell division and proliferation, potentially providing a better understanding of cancer progression.
(2) Studying gene interactions and signaling pathways: Many biological processes involve complex networks of genes that interact to perform specific functions. By knocking down a gene of interest and observing the effects on the expression of other genes, researchers can gain insight into the complex network of gene interactions. This knowledge is critical to understanding disease mechanisms and developing more effective treatments.
(3) Target validation and drug discovery: Gene knockdown cell lines can help determine whether a target gene is critical for disease progression or cancer cell survival. By successfully knocking down the gene and observing the effects on cell viability or disease-related phenotypes, researchers can evaluate the potential efficacy of drugs targeting that specific gene.
(4) Designing diseases models: Before studying disease mechanisms or testing potential therapeutic interventions, it is critical to develop valid disease models. Gene knockdown cells allow researchers to determine whether altering the expression of specific genes can reproduce disease-related phenotypes observed in patients or animal models.
Case Study 1: HeLa-Human-PARP1-Knockdown Cell Line
Figure 1. The relative expression level of Human-PARP1 was detected by Real-Time qPCR.
Case Study 2: HeLa-Human-PARP2 Knockdown Cell Line
Figure 2. The relative expression level of Human-PARP2 was detected by Real-Time qPCR.
Case Study 3: MCF7-Human-ROCK1 Knockdown Cell Line
Figure 3. The relative expression level of Human-ROCK1 was detected by Real-Time qPCR.
Case Study 4: MCF7-Human-ROCK2 Knockdown Cell Line
Figure 4. The relative expression level of Human-ROCK2 was detected by Real-Time qPCR.
Case Study 5: THP1-Human-MERTK Knockdown Cell Line
Figure 5. The relative expression level of Human-MERTK was detected by Real-Time qPCR.
Q: What is gene knockdown?
A: Gene knockdown is a technique used to reduce or suppress the expression of a specific gene in living organisms, typically through the use of short interfering RNA (siRNA) or antisense oligonucleotides.
Q: How does siRNA knockdown work?
A: siRNA knockdown, also known as RNA interference (RNAi), is a biological process that regulates the expression of specific genes by using small interfering RNA (siRNA) molecules to inhibit their activity. These RNA pairs bind to a cellular enzyme called the RNA-induced silencing complex (RISC), which uses one strand of siRNA to bind to a single-stranded RNA molecule of complementary sequence, known as mRNA. The nuclease activity of RISC then degrades the mRNA, thereby silencing the expression of the viral gene.
Q: How does shRNA knockdown work?
A: shRNA molecules are processed within the cell to form siRNA which in turn knock down gene expression. The benefit of shRNA is that they can be incorporated into plasmid vectors and integrated into genomic DNA for longer-term or stable expression, thereby knocking down target mRNAs for longer periods.
Q: What delivery methods are commonly used for siRNA and shRNA?
A: Synthetic siRNAs are introduced into cells directly, while shRNAs are often delivered using viral vectors.
Q: How long does siRNA-mediated knockdown typically last?
A: siRNA-mediated knockdown is transient and lasts for a few days to a week.
Q: What are some potential off-target effects of siRNA?
A: Due to their short length and perfect complementarity to the target mRNA, siRNAs may exhibit more off-target effects, potentially leading to unintended silencing of genes with partially similar sequences.
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