siRNA / RNAi /miRNA transfection Rat C6

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.

Get tips on using ON-TARGETplus Rat Smad2 (29357) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad2

Get tips on using ON-TARGETplus Rat Smad3 (25631) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - IEC-6 Smad3

Get tips on using ON-TARGETplus Rat Hspa5 (25617) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - AR42J Grp78/Hspa5

Get tips on using ON-TARGETplus Rat Pld2 (25097) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - RBL-2H3 Pld2

Get tips on using ON-TARGETplus Rat Atg7 (312647) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Atg7

Get tips on using ON-TARGETplus Rat Becn1 (114558) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - UMR‐106 Becn1

Get tips on using ON-TARGETplus Rat Arhgef12 (367072) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Rat - MTLn3 Larg/Arhgef12