crispr-human-repression-hpv-16-e7

Get tips on using pLV-EF1a-IRES-Puro to perform CRISPR Mouse - Deletion L929 Ppm1b

Get tips on using pCas-Guide-EF1a-GFP to perform CRISPR Mouse - Deletion αT3 Stim2

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.

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.

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

Get tips on using pX333-sgNF1-sgTrp53-Cas9 to perform CRISPR Mouse - Deletion 3T3-L1 Trp53

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 pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion AR42J FICD

Get tips on using pSpCas9(BB)-2A-GFP (PX458) to perform CRISPR Rat - Deletion AR42J Atg12