Microarray Gene expression arrays Rat chorid plexus

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.

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

Get tips on using Ccl2 siRNA to perform siRNA / miRNA gene silencing Rat - Neuronal cells MCP-1

Get tips on using HuR siRNA to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR Lipid

Get tips on using Rn_Pdcd8_1 FlexiTube siRNA to perform siRNA / miRNA gene silencing Rat - H9c2 AIF/Pdcd8

Get tips on using MicroRNA isolation kit to perform siRNA / miRNA gene silencing Rat - IEC-6 HuR

Get tips on using siRNA(r) NM_012820 Acsl1 to perform siRNA / miRNA gene silencing Rat - 3Y1 Acsl1

Get tips on using siRNA(r) NM_057107 Acsl3 to perform siRNA / miRNA gene silencing Rat - 3Y1 Acsl3

Get tips on using siRNA(r) NM_053623 Acsl4 to perform siRNA / miRNA gene silencing Rat - 3Y1 Acsl4

Get tips on using connexin 43 siRNA (r) to perform siRNA / miRNA gene silencing Rat - Pericytes Cx43