Extended Data Fig. 1: Characterization of the life-span and efficiency of RIDE.
a, Western blot analysis of the life-span of CRISPR–RNP complex. Lenti-CRISPR was used as a positive control. In total, 150 ng p24 lentiviral particles were used. 293T cells were seeded 24 h before transduction at a density of 4 × 104/well into a 24-well plate. A representative result is shown from two biologically independent replicates. b, Comparison of the gene editing efficiencies of RIDE-CRISPR and LNP-delivered CRISPR/RNP at the AAVS1 locus. In total, 200 ng p24 RIDE-CRISPR and 20 pmol RNP were used. 293T cells were seeded 24 h before transduction or transfection at a density of 4 × 104/well. RNP was mixed with Lipofectamine 2000 before transfection. c, Comparison of the gene editing efficiencies of RIDE-CRISPR, mLP-CRISPR, and Lenti-CRISPR. In total, 100 ng p24 were transduced into 1 × 104 293T cells seeded 24 h before transduction. An exception was 30 ng p24 for B2M. *P = 0.0325 for AAVS1 RIDE-CRISPR versus mLP-CRISPR, ***P = 0.0006 for AAVS1 RIDE-CRISPR versus Lenti-CRISPR, ***P = 0.0005 for Vegfa RIDE-CRISPR versus mLP-CRISPR, **P = 0.0016 for NECTIN1 RIDE-CRISPR versus mLP-CRISPR, ***P < 0.0001 for NECTIN1 RIDE-CRISPR versus Lenti-CRISPR. d, Comparison of the gene editing efficiencies of RIDE and AAV at the Nectin1 site. In total, 50 ng p24 RIDE-CRISPR and 2 × 109 GC AAV8 were transduced into 2 × 104 NIH3T3 cells seeded 24 h before transduction. e, The titer comparison of RIDE and GFP-encoding integration-deficient lentiviral vector. Indel frequency was analyzed by TIDE software (b-d). Data and error bars represent mean ± SEM from three biologically independent replicates (b-e). Unpaired two-tailed Student’s t-tests, n.s., non-significant.
