7 results on '"Bhagyashree Bachhav"'
Search Results
2. SYNTHETIC BIOLOGY APPROACHES FOR MAMMALIAN CELL FACTORY ENGINEERING
- Author
-
Laura Segatori, Bhagyashree Bachhav, Jacopo de Rossi, and Carlos D. Llanos
- Abstract
The production of high-quality recombinant proteins is critical to maintaining a continuous supply of biopharmaceuticals, such as therapeutic antibodies. Engineering mammalian cell factories presents a number of limitations typically associated with proteotoxic stress induced upon aberrant accumulation of off-pathway protein folding intermediates, which eventually culminate with the induction of apoptosis. Recent progress in mammalian synthetic biology provides unique opportunities to endow cells with programmable, user-defined behaviors, thereby addressing some of the challenges of current methods. In this review, we will discuss advances in synthetic biology to design efficient strategies for biomanufacturing. more...
- Published
- 2022
- Full Text
- View/download PDF
Catalog
3. A gene signal amplifier platform for monitoring the unfolded protein response
- Author
-
Alexander L Yang, Bhagyashree Bachhav, Sahiti D Patibandla, Carlos A. Origel Marmolejo, and Laura Segatori
- Subjects
Transcription, Genetic ,Green Fluorescent Proteins ,Protein Serine-Threonine Kinases ,Biology ,Endoplasmic Reticulum ,Chromosomes ,eIF-2 Kinase ,03 medical and health sciences ,Genes, Reporter ,Endoribonucleases ,Gene expression ,Humans ,Molecular Biology ,Gene ,030304 developmental biology ,Regulation of gene expression ,0303 health sciences ,Gene Expression Profiling ,Endoplasmic reticulum ,030302 biochemistry & molecular biology ,HEK 293 cells ,Computational Biology ,Cell Biology ,Activating Transcription Factor 6 ,Cell biology ,Chromatin ,HEK293 Cells ,Protein Biosynthesis ,Unfolded Protein Response ,Unfolded protein response ,Signal transduction - Abstract
Gene expression in mammalian cells results from coordinated protein-driven processes guided by diverse mechanisms of regulation, including protein-protein interactions, protein localization, DNA modifications and chromatin rearrangement. Regulation of gene expression is particularly important in stress-response pathways. To address the need to monitor chromosomal gene expression generating a readily detectable signal output that recapitulates gene expression dynamics, we developed a gene signal amplifier platform that links transcriptional and post-translational regulation of a fluorescent output to the expression of a chromosomal target gene. We generated a multiplex reporter system for monitoring markers of the unfolded protein response, a complex signal transduction pathway that remodels gene expression in response to proteotoxic stress in the endoplasmic reticulum. By recapitulating the transcriptional and translational control mechanisms underlying the expression of a target gene with high sensitivity, this platform provides a technology for monitoring gene expression with superior sensitivity and dynamic resolution. more...
- Published
- 2020
- Full Text
- View/download PDF
4. A PLATFORM TECHNOLOGY FOR MONITORING THE UNFOLDED PROTEIN RESPONSE
- Author
-
Bhagyashree Bachhav, Carlos A. Origel Marmolejo, Yafet Arefeayne, and Laura Segatori
- Subjects
Technology ,Unfolded Protein Response ,Gene Regulatory Networks ,Endoplasmic Reticulum ,Endoplasmic Reticulum Stress ,Article - Abstract
The unfolded protein response (UPR) is a complex signal transduction pathway that remodels gene expression in response to proteotoxic stress in the endoplasmic reticulum (ER) and is linked to the development of a range of diseases, including Alzheimer's disease, diabetes, and several types of cancer. UPR induction is typically monitored by measuring the expression level of UPR marker genes. Most tools for quantifying gene expression, including DNA microarrays and quantitative PCR with reverse transcription (RT-PCR), produce snapshots of the cell transcriptome, but are not ideal for measurements requiring temporal resolution of gene expression dynamics. Reporter assays for indirect detection of the UPR typically rely on extrachromosomal expression of reporters under the control of minimal or synthetic regulatory sequences that do not recapitulate the native chromosomal context of the UPR target genes. To address the need for tools to monitor chromosomal gene expression that recapitulate gene expression dynamics from the native chromosomal context and generate a readily detectable signal output, we developed a gene signal amplifier platform that links transcriptional and post-translational regulation of a fluorescent output to the expression of a chromosomal gene marker of the UPR. The platform is based on a genetic circuit that amplifies the output signal with high sensitivity and dynamic resolution and is implemented through chromosomal integration of the gene encoding the main control element of the genetic circuit to link its expression to that of the target gene, thereby generating a platform that can be easily adapted to monitor any UPR target through integration of the main control element at the appropriate chromosomal locus. By recapitulating the transcriptional and translational control mechanisms underlying the expression of UPR targets with high sensitivity, this platform provides a novel technology for monitoring the UPR with superior sensitivity and dynamic resolution. more...
- Published
- 2022
5. A platform for post-translational spatiotemporal control of cellular proteins
- Author
-
Laura Segatori, Santiago Martinez Legaspi, Brianna E K Jayanthi, Bhagyashree Bachhav, and Zengyi Wan
- Subjects
Computer science ,AcademicSubjects/SCI00010 ,Biomedical Engineering ,Bioengineering ,Computational biology ,localization ,Biomaterials ,03 medical and health sciences ,0302 clinical medicine ,Synthetic gene ,Process information ,orthogonal protein regulation ,Cellular proteins ,030304 developmental biology ,degradation ,0303 health sciences ,Protein function ,mammalian genetic circuits ,Subcellular localization ,Agricultural and Biological Sciences (miscellaneous) ,Protein subcellular localization prediction ,nanobody ,Post translational ,030217 neurology & neurosurgery ,Biotechnology ,Research Article - Abstract
Mammalian cells process information through coordinated spatiotemporal regulation of proteins. Engineering cellular networks thus relies on efficient tools for regulating protein levels in specific subcellular compartments. To address the need to manipulate the extent and dynamics of protein localization, we developed a platform technology for the target-specific control of protein destination. This platform is based on bifunctional molecules comprising a target-specific nanobody and universal sequences determining target subcellular localization or degradation rate. We demonstrate that nanobody-mediated localization depends on the expression level of the target and the nanobody, and the extent of target subcellular localization can be regulated by combining multiple target-specific nanobodies with distinct localization or degradation sequences. We also show that this platform for nanobody-mediated target localization and degradation can be regulated transcriptionally and integrated within orthogonal genetic circuits to achieve the desired temporal control over spatial regulation of target proteins. The platform reported in this study provides an innovative tool to control protein subcellular localization, which will be useful to investigate protein function and regulate large synthetic gene circuits. more...
- Published
- 2021
6. A yeast selection system for the detection of proteasomal activation
- Author
-
Bhagyashree Bachhav, Claire D. McWhite, Wenting Zhao, and Laura Segatori
- Subjects
0301 basic medicine ,Proteasome Endopeptidase Complex ,Saccharomyces cerevisiae Proteins ,Saccharomyces cerevisiae ,Bioengineering ,Protein Engineering ,Biochemistry ,03 medical and health sciences ,Homeostasis ,URA3 ,Molecular Biology ,biology ,Cell growth ,Chemistry ,Ubiquitin ,biology.organism_classification ,Phenotype ,Yeast ,Cell biology ,Enzyme Activation ,030104 developmental biology ,Proteasome ,Proteolysis ,Degron ,Function (biology) ,Biotechnology - Abstract
The ubiquitin proteasome system (UPS) is a complex cellular machinery that catalyzes degradation of misfolded or damaged proteins and regulates turnover of native proteins in eukaryotic cells, thus playing a crucial role in maintaining protein homeostasis. The UPS has emerged as a drug target for a diverse range of diseases characterized by accumulation of misfolded or aggregated proteins. While enhancement of UPS activity is widely recognized as a promising strategy to prevent accumulation of aberrant, off-pathway protein conformations and ameliorate the phenotypes of a wide range of protein misfolding diseases, the molecular mechanisms underlying activation of proteasomal degradation are poorly characterized. We report the development of a yeast selection platform for genome-wide selection of UPS activators. We engineered the Saccharomyces cerevisiae selection marker orotidine-5'-phosphate decarboxylase (URA3) to function as a substrate of proteasomal degradation through fusion to UPS-sensitive tags. The resulting UPS-sensitive URA3 variant links UPS activity to cell growth. The yeast selection platform reported in this study will open the way to high-throughput, genome-wide studies aimed at identifying modulators of UPS function that might provide novel target for therapeutic applications. more...
- Published
- 2018
7. Hysteretic Genetic Circuit for Detection of Proteasomal Degradation in Mammalian Cells
- Author
-
Zeng, Yimeng, primary, Bhagyashree, Bachhav, additional, Zhao, Wenting, additional, Nguyen, Tram, additional, and Segatori, Laura, additional
- Published
- 2019
- Full Text
- View/download PDF
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.