Products/Services Used | Details | Operation |
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Gene Synthesis> | The catalytic domain of VAV2 (residues 190–374, synthesized by Genscript) was cloned with BCL-xL, BH3 peptide, and linker sequences into a pDest-527 bacterial protein expression vector using Gateway recombination cloning technology by Life Technologies Human Rac1 (residues 1–177) was amplified from the original pcDNA3-EGFP-Rac1(wt) plasmid (Addgene plasmid 12980). Free peptides were synthesized by GenScript. | Get A Quote |
Bacterial Expression System> | Get A Quote |
Synthetic protein switches controlled with user-defined inputs are powerful tools for studying and controlling dynamic cellular processes. To date, these approaches have relied primarily on intermolecular regulation. Here we report a computationally guided framework for engineering intramolecular regulation of protein function. We utilize this framework to develop chemically inducible activator of RAS (CIAR), a single-component RAS rheostat that directly activates endogenous RAS in response to a small molecule. Using CIAR, we show that direct RAS activation elicits markedly different RAS-ERK signaling dynamics from growth factor stimulation, and that these dynamics differ among cell types. We also found... More