Balaji Rao
Bio
Proteins mediate and control several cellular processes. For instance, cytokine binding to cell surface receptor can trigger a series of biochemical reactions leading to outcomes such as proliferation, differentiation or death. Aberrant protein-mediated processes are involved in pathological states such as cancer. Our central hypothesis is that manipulating molecular interactions through engineered proteins can be used to understand and ultimately control cellular processes.
We have the capability to engineer biophysical properties of proteins such as binding affinity and thermal stability as well as soluble protein expression. We use powerful protein engineering tools such as yeast surface display and mRNA display to generate proteins with desired properties. These proteins enable us to study and ultimately control cellular processes and may potentially be clinically relevant.
We are particularly interested in the area of stem cell bioengineering. Stem cells are “master” cells that can self-renew as well as develop into many different cell types. Stem cells have great potential in regenerative medicine. They can also be used as a basis to develop model systems for drug evaluation. One of the major challenges in this area is to understand and control the molecular decisions that control stem cell fate. Our approach involves the use of engineered proteins to quantitatively study and ultimately control molecular interactions that govern stem cell fate.
Research Focus Areas: Molecular and Cell Bioengineering, Molecular Control of Cellular Processes, Stem Cell Bioengineering.
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Publications
- Laminin switches terminal differentiation fate of human trophoblast stem cells under chemically defined culture conditions , JOURNAL OF BIOLOGICAL CHEMISTRY (2023)
- Experimental and Analytical Framework for "Mix-and-Read" Assays Based on Split Luciferase , ACS OMEGA (2022)
- Experimental and Analytical Framework for "Mix-and-Read" Assays Based on Split Luciferase , ACS OMEGA (2022)
- Modified Histone Peptides Linked to Magnetic Beads Reduce Binding Specificity , INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022)
- Mapping the residue specificities of epigenome enzymes by yeast surface display , CELL CHEMICAL BIOLOGY (2021)
- Quantitative Yeast-Yeast Two Hybrid for the Discovery and Binding Affinity Estimation of Protein-Protein Interactions , ACS SYNTHETIC BIOLOGY (2021)
- Screening of Yeast Display Libraries of Enzymatically Treated Peptides to Discover Macrocyclic Peptide Ligands , INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2021)
- Two distinct trophectoderm lineage stem cells from human pluripotent stem cells , JOURNAL OF BIOLOGICAL CHEMISTRY (2021)
- A Fyn biosensor reveals pulsatile, spatially localized kinase activity and signaling crosstalk in live mammalian cells , ELIFE (2020)
- Discovery of Membrane-Permeating Cyclic Peptides via mRNA Display , BIOCONJUGATE CHEMISTRY (2020)
Grants
We propose to develop peptide-based adsorbents for the purification of AAVs to be used for gene therapy.
The goal of this project is to develop chemically defined synthetic matrices to conduct in vitro studies on trophoblast differentiation in 3D cultures.
There is substantial evidence that chromatin (genomic DNA and the proteins, RNAs, and chemical motifs bound to it) is a central regulator of diverse cellular and disease processes, in particular through its regulation of gene expression. Despite the wide-spread acceptance of its importance and relevance throughout biology, it is remarkable that our understanding of chromatin??????????????????s mechanisms and functions remains very limited and reliant on largely correlative observations or non-specific genome-wide perturbations. Here we propose the engineering of two synergistic and parallel sets of tools. One set of tools will sense and report on the dynamic biochemical and conformational states of chromatin in living mammalian cells. The other set of tools will dynamically induce changes in or ????????????????edit??????????????? chromatin biochemistry and conformation at specific user-defined genomic loci.
The goal of this project is to study formation of the trophectoderm layer of the human embryo using in vitro models derived from human and nonhuman primate pluripotent stem cells.
The goal of the NCSU subcontract is to investigate the effect of nicotinamide and endothelin-1 on trophoblast differentiation and invasion, in an in vitro model of human trophoblast development.
We propose to establish a microfluidics-enabled in vitro system to quantitatively interrogate the effects of external stimuli (BPA, oxygen concentration/gradient) and intercellular commu-nication (trophoblast-macrophage interactions) on trophoblast differentiation and invasion in 3D cultures.
The objective of this proposal is to obtain funding from the North Carolina Biotechnology Center to purchase a unique, automated, and high throughput slide scanning imager manufactured by Olympus. The new equipment allows for rapid acquisition of highly detailed images of cells and tissues. In biological research, often the generation of large amounts of samples from experiments is desired to tackle important research questions. Yet, sample generation often is not the limiting factor. Rather the imaging and data analysis of large numbers of samples is usually prohibitive and limits the types and impact of research questions that can be asked. As described in this proposal, the Olympus imaging system would unlock a wide range of important biomedical questions in diverse systems from zebrafish to human, bone marrow to brain, by providing an over 10-fold improvement in sample imaging throughput and analysis. The instrument will be installed within an NCSU Shared Core Research Facility, the Cellular and Molecular Imaging Facility (CMIF), which lacks the proposed technology currently, and is staffed by professional staff that will manage, maintain, and operate the proposed equipment. Acquisition of the Olympus slide scanner and installation within CMIF will increase the rate of data acquisition by research groups at NCSU and, in so doing, ensure their continued competitiveness for research funding.
The goal of this project is to investigate placental signals to the brain during development, using in vitro models of the placenta and brain.
The goal of this project is to validate a defined culture system for generation of trophoblast cells from human embryonic stem cells.
The goal of this proposed project is to develop protein reagents that can be used to isolate specific cellular subtypes non-destructively from heterogeneous samples.