RNA Interference: Features, Application and Perspectives

Author: Bennie George

RNA interference (RNAi) technology makes it possible to effectively regulate gene expression. The application of this technology in the laboratory allows scientists to have a glimpse of the complex biological systems of humans and mammals. The ability of RNAi to knock down target genes has allowed scientists to identify and validate drug targets, as well as discover cellular signaling pathways that are critical to biological survival and disease control. This technology is essential for the development of new therapeutic drugs and therapies.

As people discover all aspects of the RNAi mechanism, the mechanism of RNAi becomes more and more clear. In the past few years, important insights have been gained in elucidating the mechanism of RNAi. The combination of results obtained from several in vivo and in vitro experiments has been gelatinized into a two-step mechanism model of RNAi/PTGS. The first step, named the RNAi initiation step, involves combining RNA nuclease with large dsRNA and cutting it into discrete RNA fragments (siRNA) of about 21 to 25 nucleotides. In the second step, these siRNAs are added to the polynucleotide enzyme complex RISC, which degrades homologous single-stranded mRNA. At present, little is known about RNAi intermediates, RNA-protein complexes, and the formation mechanisms of different complexes during RNAi.

In addition to being an intensive, early-stage basic research area, RNAi processes are also key to future technology applications. The genome sequencing project generated a lot of information. However, the ultimate goal of such projects is to speed up the identification of genetic biological functions. The function of genes can be analyzed by appropriate analysis methods by examining the phenotype of organisms that contain mutations in the genes, or based on the knowledge gained from studies of genes related to other organisms. However, a large part of the genes identified by the sequencing project are new, and functions cannot be quickly assigned by these conventional methods.

In view of the fact that RNAi is easy to apply, whole-genome screening by RNAi may become a common selection method in the near future. RNAi can facilitate drug screening and development by identifying genes that can confer drug resistance or can improve its mutant phenotype by drug treatment, and provide information about the mode of action of new compounds. Although RNAi is unlikely to replace existing gene knockout technology, it may have a huge impact on organisms that are not suitable for gene knockout strategies. The method of studying the simultaneous function of many similar genes in an organism may also be a selection method. In these organisms, there is redundancy for specific functions, because many of these genes can be silenced at the same time.

Considering the gene-specific characteristics of RNAi, it is conceivable that this method will play an important role in therapeutic applications. Because siRNAs guide cellular RNAi biology, they are potential therapeutic agents because they can down-regulate the expression pattern of mutated genes in diseased cells. However, the central assumption of this hypothesis is that the effect of exogenous siRNA application will still be gene-specific and will not show non-specific side effects related to off-target hybridization with mismatches, protein-nucleic acid binding, etc. Not even one nucleotide in 19 to 20 molecules of siRNA can effectively destroy the appropriate degradation of target mRNA, and it is necessary to confirm the gene specificity of siRNA in the whole genome.

With an in-depth understanding of the mechanism of RNAi, more effective methods of gene function analysis may be developed. We may learn more about geriatric diseases, neurological diseases, genetic imprinting, and nuclear superiority in plants, so these processes may be controlled in the future. At the same time, by designing better plasmid- or virus-based vectors, siRNAs can be delivered to the appropriate tissues at the appropriate time, and this knockout technique may be greatly improved, thereby bring a new look to therapeutic gene silencing.

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