CRISPR Human Deletion

Get tips on using pcDNA-mC/EBPb to perform CRISPR Mouse - Activation 3T3-L1 C/EBPβ

Get tips on using lenti sgRNA(MS2)_zeo backbone to perform CRISPR Mouse - Activation C2C12 FST

Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Activation C2C12 Lama1

Get tips on using pMSCV-LTR-dCas9-VP64-BFP to perform CRISPR Mouse - Activation Neuro-2a Sim1

Get tips on using pX330-U6-Chimeric_BB-CBh-hSpCas9 to perform CRISPR Mouse - Activation 3T3-L1 eIF5A1

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Mouse - Activation 3T3-L1 ZPR9

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

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

Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Mouse - Activation 3T3-L1 Adrb3

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.