This repository hosts the data, processing, analysis and plotting for the jAspSnFR3 manuscript titled:
An engineered biosensor enables dynamic aspartate measurements in living cells
The sensor was tested by stable expression (lentivirus infection, monoclonal cell line) in different cell lines (H1299, HT1080 and HEK293T) and live cell imaged (GFP/RFP signal monitoring on Incucyte S3). The sensor signal was captured using the GFP channel and normalized to RFP channel signal, using either RFP fused to the sensor, or a nuclear localized RFP.
Experiments were performed as either "temporal" (Figure 2 in manuscript) or "steady-state" (Figure 3 in manuscript). With temporal experiments tracking sensor signal over time after a pertubation, and steady-state experiments correlating sensor signal with LCMS determined aspartate concentration after a pertubation has reached a near steady-state level of sensor signal.
Temporal data is read, analyzed and plotted here:
temporal_signal.ipynb
Steady-state data is read, analyzed and plotted here:
steady-state_conc.ipynb
For LCMS based amino acid quantification, isotope dilution was used and calibration curves generated. These calibration curves are plotted here:
AA_calibration-curves.pdf