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. 2020 Aug 21;6(9):1564–1571. doi: 10.1021/acscentsci.0c00940

Figure 4.

Figure 4

Evaluation of tyrosine–cysteine bioconjugates in cell culture assays. (a) Evaluation of modified CRISPR-Cas9 ribonucleoproteins (RNPs) for gene editing in neural progenitor cells. The assay measured the increase in fluorescent protein expression following successful editing at a tdTomato locus. Cas9 modified with two copies of the SV40 nuclear localization sequence (Cas9-2SV40) showed a 20-fold increase in editing efficiency compared with unmodified Cas9. Data were collected 72 h after treatment. Error bars show standard deviations of triplicates. (b) The coupling reaction was successful for the coupling of GFP Y200C to a HER2-binding scFv-GGY construct, as measured by ESI-TOF. The observed product mass was consistent with the linkage depicted in (c). (d) Flow cytometry analysis of SK-BR-3 (HER2+) and MDA-MB-468 (HER2−) breast cancer cells treated with the trastuzumab scFv–sfGFP construct showed HER2-specific cell binding. Gating and statistics are shown in Figure S18.