Fig. 5: EVs mediate functional delivery of Cas9 in primary human T cells.

a, Illustration of function delivery evaluation. 2.0×10¹⁰ EVs were incubated with 5.0×10⁴ CD4⁺ T cells for 6 d prior to genomic DNA extraction and HTS analysis. b, Frequency of indels detected at the Cas9-targeted CXCR4 locus. The sgRNA recognition site (green), PAM sequence (underlined, red), and predicted cut site (amplicon position 26, scissors) are shown. Total percentage of HTS reads classified as “edited” represents the area under the histogram trace shown for each sample. c, Distributions of EV-Cas9-mediated edits, by type. DNA amplicon position is plotted on the abscissa and length of the edit observed is plotted on the ordinate, while the size of each dot scales with the number of edits that meet that description. Each read is uniquely classified as a deletion, insertion, or substitution such that no one read contributes to more than one dot in this panel. In the case of substitutions, the positive ordinate reports the insertion portion of the edit, and the negative ordinate reports the deletion portion of the edit such that each edit is represented by two dots. In this panel, deletions are reported by placing a dot at the midpoint of the deleted segment. To help explain the apparent “V” pattern, dots are colored blue to indicate cases where one end of the deleted segment corresponds to the predicted cut region, presumably corresponding to a subset of the DNA repair outcomes observed. Sample dot coloring is as in b.