| RNA interference (RNAi) is the process of mRNA | | | | miRNAs were discovered through their critical roles in |
| degradation that is induced by double-stranded RNA | | | | development and cellular regulation, and represent a |
| in a sequence-specific manner. RNAi has been | | | | large class of evolutionarily conserved RNAs. miRNAs |
| observed in all eukaryotes, from yeast to mammals. | | | | have always been recognized as being of |
| The RNAi pathway is thought to be an ancient | | | | endogenous origin. RNA interference has emerged as |
| mechanism for protecting the host and its genome | | | | a natural mechanism for silencing gene expression |
| against viruses and rogue genetic elements that use | | | | over the past decade. This ancient cellular antiviral |
| double-stranded RNA (dsRNA) in their life cycles. | | | | response can be harnessed to allow specific inhibition |
| They have also been shown to play a role not only in | | | | of the function of any chosen target genes, including |
| mRNA and dsRNA stability/degradation, but also in | | | | those involved in causing diseases such as cancer, |
| regulation of translation, transcription, chromatin | | | | AIDS, and hepatitis. It is already proving to be an |
| structure, and genome integrity. In plants and animals, | | | | invaluable research tool, allowing much more rapid |
| RNA silencing has been adapted to play a critical role | | | | characterization of the function of known genes. |
| in regulation of cell growth and differentiation using a | | | | More importantly, the technology considerably |
| class of small RNAs. In the RNA interference process, | | | | bolsters functional genomics to aid in the identification |
| the dsRNAs get processed into 20-25 nucleotide (nt) | | | | of novel genes involved in disease processes. Last |
| small RNAs by an RNase III-like enzyme called Dicer. | | | | but not the least the technology can be harnessed |
| Then, the small RNAs assemble into | | | | as a novel therapeutic agent and is suitable for |
| endoribonuclease-containing complexes known as | | | | combating viral diseases, cancers and inflammatory |
| RNA-induced silencing complexes (RISCs), unwinding | | | | diseases. |
| in the process. The small RNA strands subsequently | | | | Imgenex (San Diego) recently launched the |
| guide the RISCs to complementary RNA molecules, | | | | pSuppressorAdeno construction kit for adenovirus |
| where they cleave and destroy the cognate RNA | | | | mediated gene knockdown. The kit provides the |
| (effecter step). Cleavage of cognate RNA takes | | | | ability to infect a broad range of cell types, including |
| place near the middle of the region bound by the | | | | many primary cell lines as well as dividing and |
| siRNA strand. The small RNAs that provide target | | | | nondividing cells, according to a company official. The |
| specificity to the silencing machinery includes short | | | | kit also offers the flexibility to validate sequences |
| interfering RNAs (siRNAs), repeat-associated siRNAs | | | | using the nonviral expression plasmid prior to |
| (rasiRNAs), and microRNAs (miRNAs) and is | | | | construction of adenoviruses, notes Sujay K. Singh, |
| distinguished by their origin. siRNAs are processed | | | | Ph.D., president and CEO of Imgenex, which markets |
| from dsRNA precursors made up of two distinct | | | | plasmid-based RNA interference (RNAi) products. |
| strands of perfectly base-paired RNA, while miRNAs | | | | “One of the greatest advantages is the ability of |
| originate from a single, long transcript that forms | | | | recombinant adenovirus vectors to reduce gene |
| imperfectly base-paired hairpin structures. siRNAs | | | | expression both in vitro and in vivo,” he adds. |
| were originally identified as intermediates in the RNAi | | | | RNAi, initially considered a bizarre attribute of petunias |
| pathway after induction by exogenous dsRNA; | | | | and later a gene-silencing mechanism in worms, is |
| however, endogenous sources of siRNAs have now | | | | creating a stir as one of the hottest new |
| been recognized. The endogenous siRNAs are derived | | | | technologies in molecular biology. It is revolutionizing |
| from repetitive sequences within the genome, and | | | | the field of functional genomics. |
| are termed repeat-associated siRNAs, or rasiRNAs. | | | | |