Figure 1: TRAC and PD-1 knock-out efficiency in human primary T cells.
Human primary T cells were cultured for the test. The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) or GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP (Z03624) or wild type Cas9, and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services ) for human TRAC and PD-1 gene knock-out by electroporation. After transfection and cell culture, the TRAC knock-out efficiency was measured by FACS while PD-1 knock-out efficiency was measured by Sanger sequencing. Both GenScript Ultra SpCas9 and eSpCas9 show high editing efficiency.
Note: This experiment used the following:
Cas9: sgRNA (molar ratio) =1:2
RNP amount: ~ 25 pmol
Cell number: 1.0 × 10⁶ cells

Figure 4: Gene knock-in efficiency at TRAC site in primary T cells.
The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) or GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP (Z03624), sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) and dsDNA HDR template (synthesized by GenScript Double-strand DNA Synthesis Services) for gene knock-in at TRAC site in primary T cells by electroporation. After transfection and cell culture, the gene knock-in efficiency was analyzed by FACS. Both GenScript GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) and GenCRISPR™ Ultra eSpCas9-2NLS-basic GMP (Z03624) show high gene knock-in efficiency.
Note: This experiment used the following:
SpCas9/eSpCas9: sgRNA (molar ratio) =1:2
RNP amount: ~ 25 pmol
Cell number: 1.0 × 10⁶ cells

Figure 3: RAB11A knock-out efficiency in 293T cells.
Human 293T cells were cultured for the test. The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) or wild type Cas9 proteins from competitors and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for human RAB11A gene knock-out by electroporation. After transfection and cell culture, the gene editing efficiency was analyzed by Sanger sequencing. GenScript GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) shows higher editing efficiency.
Note: This experiment used the following:
SpCas9: sgRNA (molar ratio) =1:3
RNP amount: 7.5 pmol
Cell number: 2.0 × 10⁵ cells

Figure 2: CD96 and NKG2A knock-out efficiency in NK92 cells.
Human NK92 cells were cultured for the test. The cells were transfected with GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) and sgRNA (synthesized by GenScript CRISPR Synthetic sgRNA Services) for human CD96 or NKG2A gene knock-out by electroporation. After transfection and cell culture, the gene editing efficiency were analyzed by Sanger sequencing. GenScript GenCRISPR™ Ultra NLS-SpCas9-basic GMP (Z03623) shows high editing efficiency.
Note: This experiment used the following:
SpCas9: sgRNA (molar ratio) = 1:1.2
RNP amount: 80 pmol
Cell number: 4.0 × 10⁵ cells