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 type Source Media Cell seeding density/well (× 103) one day before transfection TrueCut Cas9 Protein v2/gRNA (ng/pmoles) Lipofectamine Cas9 Plus Reagent/well (μL) Lipofectamine CRISPRMAX Reagent/well (μL)
Well format 96-well 24-well 6-well 96-well 24-well 6-well 96-well 24-well 6-well 96-well 24-well 6-well
HEK293 Human embryonic kidney DMEM 18 90 450 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.4 2 10
U2OS Human osteosarcoma McCoy5A 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
A549 Human epithelial lung carcinoma DMEM 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
THP1 Human peripheral blood monocyte leukemia RPMI 10 50 250 400/2.4 2000/12 10000/60 0.8 4 20 0.3 1.5 7.5
K562* Human leukemia bone marrow RPMI 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
iPSC* Human induced pluripotent stem cells Essential 8 8 40 200 300/2 1500/10 7500/50 0.6 3 15 0.3 1.5 7.5
HepG2 Human hepatocellular carcinoma DMEM 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
MDA- MB231 Human epithelial (breast) adenocarcinoma DMEM 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
N2A Mouse brain neuroblastoma DMEM 10 50 250 250/1.5 1250/7.5 6250/37.5 0.5 2.5 12.5 0.3 1.5 7.5
*Use the Neon Transfection System for higher editing efficiency.