ChIP H3K27me3 Human Goat

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.

Get tips on using MAGnify™ Chromatin Immunoprecipitation System to perform ChIP Rat - Spinal cord

Get tips on using MAGnify™ Chromatin Immunoprecipitation System to perform ChIP Mouse - Cardiac fibroblasts

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Mouse - CD4+ T

Get tips on using Imprint® Chromatin Immunoprecipitation Kit to perform ChIP Rat - Brain microvessels

Get tips on using truChIP Chromatin Shearing Kit with Formaldehyde to perform ChIP Mouse - Brain

Get tips on using Human ShhN ELISA to perform ELISA Human - ShhN

Get tips on using Bcl-11B (D6F1) XP® Rabbit mAb #12120 to perform ChIP Anti-bodies CtIP/BCL11A

Get tips on using Human ANGPTL3 ELISA to perform ELISA Human - Angiopoietin-Like 3 (AngptL3)

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.