Extended Data Fig. 9: Validation the stability of on-target editing.
From: dCas9-based gene editing for cleavage-free genomic knock-in of long sequences
(a) Workflow of the long-term time-course experiments to evaluate the editing outcome stability using dCas9–SSAP editor. (b) Flow cytometry analysis of knock-in gene editing at HSP90AA1, ACTB endogenous loci at different time points post delivery of dCas9–SSAP and donor DNA. (c, d) Representative crystal violet staining images for the on-target puromycin knock-in at HSP90AA1 and ACTB locus. Scale bar = 500 um. The assay has been performed 3 times with similar results. (e) Quantify HSP90AA1 and ACTB gene expression levels in HEK293T cells by bulk RNA-seq analysis, demonstrating notable higher levels of HSP90AA1 expression. This led to the better cell survival in the HSP90AA1 group compared with ACTB group. Data from 2 biologically independent experiments are presented.
