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[Preprint]. 2024 Feb 4:2024.01.31.578281. [Version 1] doi: 10.1101/2024.01.31.578281

Figure 1: A mAbCII-programmed synNotch receptor licenses cells for potent and specific responses to type II collagen.

Figure 1:

(A) The type II collagen (CII)-sensitive antibody mAbCII serves as a synNotch recognition motif, rendering a new synNotch receptor, CII-synNotch. CII-synNotch facilitates cell recognition of CII fibers, leading to transmembrane cleavage of a synthetic transcription factor for versatile output responses. (B) CII-synNotch MSCs were plated on surfaces treated with solutions of varying concentrations of collagen I (CI) and CII (t=72hr). Red dashed line indicates basal signal from untreated cells. (C) Luminescence values of engineered cells on either 25 µg/ml CI or CII (t=72hr). (D) Fluorescence microscopy of constitutively iRFP+ CII-synNotch MSCs with inducible mCherry expression (t=72hr). (E) Activation of synNotch MSCs with either a mAbCII receptor or non-cognate (anti-GFP) receptor, measured via luciferase detection (t=72hr). Scale = 200 µm. Data: n=3 replicates, plotted as mean ± SEM. Statistical analysis: one-way ANOVA with Tukey’s post-hoc analysis, ****p < 0.0001. NT: No treatment.