Cell-Line Specific CRISPR RNP Transfection Conditions Using Lipofectamine CRISPRMAX Reagent

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.

For Research Use Only. Not for use in diagnostic procedures.