RNA interference is a recently discovered functional tool. When RNA is introduced into a cell, it will eventually cause the degradation of complementary mRNA in the cell, resulting in the blocking of gene functional activity.
PTGS posttranscriptional gene silencing (posttranscriptional gene silencing); a phenomenon that is first identified in plants and then found in animals. Although PTGS was originally described as an endogenous method of viral defense and transposon silencing, it has now become a new and exciting research tool-RNAi interference.
Cosuppression (Cosuppression) is a special case of gene silencing, where the RNA derived from the transgene and the homologous endogenous gene are simultaneously suppressed.
Quielling is used to describe synergistic co-suppression in pink bread mold (Neurosporacrassa). This term is only used to describe silence in fungi.
dsRNA Double-stranded RNA (Double-stranded RNA); usually refers to long or full-length RNA double-stranded. These large dsRNAs fully initiate the shutdown of host cells in most mammalian cell types; then the cells begin to reduce the expression of non-target genes and eventually undergo apoptosis.
siRNA short (or small) interfering RNA (Short (or small) interfering RNA); mammalian cells are involved in short RNA double strands of 21-23 nucleotides that cause an RNAi response.
RISCRNA-induced silencing complex (RNA-inducedsilencingcomplex); that is, the presumed multi-protein complex that combines siRNA and intracellular mRNA, activates the cleavage mechanism (probably endonucleotide), releases mRNA and degrades it.
shRNA short hairpin RNA (ShorthairpinRNA); also known as short interfering hairpin (shortinterferinghairpin); in vivo through RNAi to reduce the expression of complementary sequence genes.
PolIII promoter PolIII promoter U6 or HI (PolIIIpromoter, U6orHI) is usually used to drive shRNA production. The PolIII promoter has all the elements necessary to initiate transcription upstream of a certain start site.
PolIII terminator PolIII terminator (PolIIIterminator); 4 or more T, provides PolIII-dependent transcription termination.
U6 promoter U6 promoter (U6promoter) is a polIII type promoter that allows the production of small RNAs with defined ends (see polIII promoter).
The entry vector contains an attL site and is a Gateway® vector used to clone DNA fragments. The U6 entry vector, pENTR® / U6, is specifically designed to use the U6polIII type promoter and terminator sequences. After the RNAi target sequence is cloned, it will form shRNA in mammalian cells.
Destination vector The destination vector (DEST) suitable for Gateway® contains attR sites, which can be recombined with entry clones to construct expression vectors. The BLOCK-iT ™ RNAi Lentivirus and Adenovirus DEST vectors accept the RNAi box from the U6 entry vector. In addition, there are DEST vectors for expressing natural proteins or fusion proteins in prokaryotic, yeast, insect or mammalian systems.
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