rna-isolation-purification-tissue-rat-pituitary-gland

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

RNAi or RNA interference is a common method to suppress gene expression in vitro/in vivo by utilizing the inherent microRNA machinery, without introducing a total gene knockout. 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.

Get tips on using Rat TGF Beta 1 PicoKine™ ELISA Kit to perform ELISA Rat - TGF-beta 1

Get tips on using Rat Superoxide dismutase [Mn], mitochondrial(SOD2) ELISA kit to perform ELISA Rat - SOD2/Mn-SOD

Get tips on using Rat IL-1 Beta PicoKine™ ELISA Kit to perform ELISA Rat - IL-1 beta

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized RMA-S

Get tips on using Active rat PAI-1 functional assay ELISA kit to perform ELISA Rat - Serpin E1/PAI-1

Get tips on using Rat plasminogen activator inhibitor 1,PAI1 ELISA Kit to perform ELISA Rat - Serpin E1/PAI-1

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - immortalized RMA-S

Get tips on using Pierce™ Cell Surface Protein Isolation Kit to perform Protein isolation Tissue - Human aortic endothelial cells