Fig. 5: Type V-K CAST are broadly applicable for the integration of a therapeutically relevant, gene-sized donor.
From: Integration of therapeutic cargo into the human genome with programmable type V-K CAST
A On-target integration efficiency of type V-K CAST in K562 cells at AAVS1 target T1 (n = 3). B Integration efficiencies in Hep3b at AAVS1 target T1 at varying doses of an all-in-one helper plasmid (AIO), donor plasmid, and ClpX plasmid concentrations (Supplementary Data 3) (n = 2). C Integration efficiencies determined by NGS when delivered with a linear donor at AAVS1 target T1 (single experiment). D Schematic of Factor IX (FIX) donor plasmids. E On target efficiencies of FIX donor with varying ratios of CAST AIO expression plasmid and FIX donor plasmids. F Relative efficiencies with FIX pDonor plasmid with and without multiple copies of the sgRNA cassette. Fold increases were normalized to the mNeon donor plasmid control with one copy of the sgRNA. G Relative rates of LE and RE junctions normalized to the LE from FIX integration experiments. Quantified integration efficiencies in A–C, E–G are shown as the mean (bar) of biological replicates (dots) with one standard deviation (whisker). Null represents the non-targeting spacer (negative control) and T1 represents the AAVS1 target 1. Source data are provided as a Source Data file.
