The following cell line-specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Lipofectamine CRISPRMAX Transfection Reagent. Further optimization of the transfection conditions may be necessary for best results.

NOTE: Scroll to the right within the table to see all data.

Cell typeSourceMediaCell seeding density/well (× 103) one day before transfectionTrueCut Cas9 Protein v2/gRNA (ng/pmoles)Lipofectamine Cas9 Plus Reagent/well (μL)Lipofectamine CRISPRMAX Reagent/well (μL)
Well format96-well24-well6-well96-well24-well6-well96-well24-well6-well96-well24-well6-well
HEK293Human embryonic kidneyDMEM1890450250/1.51250/7.56250/37.50.52.512.50.4210
U2OSHuman osteosarcomaMcCoy5A1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
A549Human epithelial lung carcinomaDMEM1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
THP1Human peripheral blood monocyte leukemiaRPMI1050250400/2.42000/1210000/600.84200.31.57.5
K562*Human leukemia bone marrowRPMI1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
iPSC*Human induced pluripotent stem cellsEssential 8840200300/21500/107500/500.63150.31.57.5
HepG2Human hepatocellular carcinomaDMEM1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
MDA- MB231Human epithelial (breast) adenocarcinomaDMEM1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
N2AMouse brain neuroblastomaDMEM1050250250/1.51250/7.56250/37.50.52.512.50.31.57.5
*Use the Neon Transfection System for higher editing efficiency.