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Fall 2015

Lai, Yan Tiffany

ubiquitin modifier 1 (SUMO-1) in pancreatic islet cells. The creation of these plasmids is a proof of the versatility and modularity of the ddFP and its related FP exchange (FPX) biosensor design strategy. We also report the engineering of a dark “A” copy (DA) for ddFP applications. By a simple rational

The expansion of the fluorescent protein (FP) colour palette through discovery of more species possessing FPs and by subjecting them to intensive protein engineering efforts has greatly aided the effectiveness of FP-based biosensor designs. Currently, the main FP-based methods for detecting protein

may facilitate multicolour imaging of biosensors in live cells due to the fact that they have a narrower spectral profile than a FRET pair. In this thesis, we describe our efforts to expand the range of applications of ddFP in live cells. We report the creation of systems for the sensing of global O

Fall 2014

Zhao, Yongxin

-based indicators remains a challenging engineering problem, mainly due to lack of structural information for rational design and effective methodologies of protein engineering. The goal of this thesis work is to tackle the long-standing challenge of engineering FP-based indicators for improved

engineering of single FP-based Ca2+ indicator and led to several variants with improved performance and various new colors. This palette of new Ca2+ indicators enables simultaneous monitoring of Ca2+ transients in different cellular compartments or different types of cells, which opens up a new era of

Fall 2020

Lu, Xiaocen

describe my efforts in development of the second generation of PhoCl variants (PhoCl2) with improved rate and efficiency of dissociation. To better understand the structural changes associated with photoconversion and peptide dissociation, we determined the X-ray crystal structures of the green state, the

red state, and the cleaved empty barrel of PhoCl1. Guided by the structural information, we further engineered PhoCl using a NanoLuc luciferase-based complementation assay for screening. Two variants of the second generation of PhoCl variants were developed via directed evolution. These variants were

nanoparticles by engineering protein hybrids composed of encapsulin and a light-sensitive protein. Encapsulin is a virus capsid-like natural nanocapsule with diameter of 25 nm to 32 nm, which is used by bacteria to isolate toxic enzymatic activities. I demonstrated in mammalian cell expression system that

Spring 2021

Wu, Sheng Yi

enable imaging in live cells with a spatiotemporal resolution and can be targeted to a specific subcellular region of a cell or a specific cell type of a tissue. While most biomolecules can be visualized with a fused fluorescent protein, more sophisticated biosensors are required to report a specific

, a DipdTFP/dTomato FRET pair was engineered by replacing the exterior residues of dTFP with that of dTomato. The surface residue engineering successfully allows heterodimer to form, but with a high affinity. More work is required to weaken this interaction for proper protein-protein interaction