Get tips on using ON-TARGETplus Rat Rhog (308875) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 RhoG
Get tips on using ON-TARGETplus Rat Trio (310192) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 Trio
Get tips on using ON-TARGETplus Rat Pak1 (29431) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 Pak1
Get tips on using ON-TARGETplus Rat Nos3 (24600) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - NPC Nos3
Get tips on using ON-TARGETplus Rat Klf15 (85497) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - NRCM Klf15
Get tips on using ON-TARGETplus Rat Atf4 (79255) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - PC12 Atf4
Get tips on using ON-TARGETplus Rat Dcp1a (361109) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - PC12 Dcp1a
Get tips on using ON-TARGETplus Rat Lsm1 (364624) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - PC12 Lsm1
Transfection is a powerful technique that enables the study of the function of genes and gene products in cells. Based on the nature of experiments, we may need a stable DNA transfection in cells for persistent gain-of-function or loss-of-function of the target gene. For stable transfection, integration of a DNA vector into the chromosome is crucial which requires selective screening and clonal isolation. By carefully selecting a viral delivery system and related reagents we can ensure safe and highly-efficient delivery of expression constructs for high-level constitutive or inducible expression in any mammalian cell type.
miRNA is the inherent gene silencing machinery which can have more than one mRNA target, whereas siRNA can be designed to target a particular mRNA target. By design, both siRNA and miRNA are 20-25 nucleotides in length. The target sequence for siRNAs is usually located within the open reading frame, between 50 and 100 nucleotides downstream of the start codon. There are two ways in which cells can be transfected with desired RNAi: 1. Direct transfection (with calcium phosphate co-precipitation or cationic lipid mediated transfection using lipofectamine or oligofectamine), and 2. Making RNAi lentiviral constructs (followed by transformation and transduction). Lentiviral constructs are time consuming, but provide a more permanent expression of RNAi in the cells, and consistent gene silencing. Direct transfection of oligonucleotides provides temporary genetic suppression. Traditional methods like calcium phosphate co-precipitation have challenges like low efficiency, poor reproducibility and cell toxicity. Whereas, cationic lipid-based transfection reagents are able to overcome these challenges, along with applicability to a large variety of eukaryotic cell lines. When using oligos, the ideal concentration lies between 10-50nM for effective transfection.
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