Cell-cell contact-induced gene editing/activation in mammalian cells using a synNotch-CRISPR/Cas9 system

摘要

. (A) Diagram of the synNotch system. (B) Flow cytometry analyses showed the activation levels of mCherry in the new synNotch systems. The receiver cells were U2OS cells with a mCherry reporter. The sender cells were K562 cells with/without CD19. Red: CD19+, black: CD19−. M: mouse Notch, H: human, Fly: drosophila, Z: zebrafish. (C) Normalized fluorescence intensity of mCherry in flow cytometry ( = 3, Student’s test, ** < 0.01, error bars, SEM). (D) Activation of EGFP in H1, Z3 and eZ3 systems. (E) Normalized fluorescence intensity of EGFP in flow cytometry ( = 3, Student’s test, ** < 0.01, error bars, SEM). For (D) and (E), the receiver cells were MCF7 cells with an EGFP reporter. (F) Western blot showed the secretion of the SIRPɑ-Fc protein in the H1, Z3 and eZ3 systems. The receiver cells were MCF7 cells with a downstream SIRPɑ-Fc