rna-sequencing-mouse-3t3-l1

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.

Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.

Get tips on using Accell Mouse Nrep (27528) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 P311/Nrep

Get tips on using hCas9 to perform CRISPR Mouse - Deletion 3T3-L1 ATP7A

Get tips on using lentiCRISPR v2 to perform CRISPR Mouse - Deletion 3T3-L1 TEAD

Get tips on using pLKO.1 puro to perform CRISPR Mouse - Deletion 3T3-L1 Epac1

Get tips on using Kdm2b siRNA to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 Fbxl10

Get tips on using Stk11 siRNA to perform siRNA / miRNA gene silencing Mouse - 3T3-L1 Stk11

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

Get tips on using APC BrdU Flow Kit to perform Cell cycle assay mouse - 3T3-L1