CRISPR Rat Deletion

Get tips on using TRIzol Reagent to perform RNA isolation / purification Tissue - Human Retina

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Tissue - Mouse Retina

Get tips on using NucleoSpin® RNA Plus to perform RNA isolation / purification Tissue - Mouse Retina

Get tips on using LAMP-1 Antibody (H4A3) to perform Autophagy assay cell type - Proximal tubular cells (rPT)

Get tips on using AllPrep DNA/RNA Mini Kit to perform RNA isolation / purification Tissue - Human Retina

Get tips on using Gibco DMEM/F-12, HEPES to perform 3D Cell Culture Media hiPSC-derived retinal organoids

Get tips on using Anti-Beclin 1 antibody (ab62557) to perform Autophagy assay cell type - Proximal tubular cells (rPT)

Get tips on using DMEM, high glucose, GlutaMAX™ Supplement, pyruvate to perform 3D Cell Culture Media hiPSC-derived retinal organoids

Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform 3D Cell Culture Media hiPSC-derived retinal organoids

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.