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Principal Investigator: STANLEY FIELDS
Organization: UNIVERSITY OF WASHINGTON
Fiscal Year: 2019
Award: $295,771
Funding agency: National Institute of General Medical Sciences
Project Summary
In TR&D 1, we develop approaches to make complex changes to cells, and we generate biosensors to report
on the effects of some of these changes. In the first aim, we establish strategies to rapidly and combinatorially
up- or down-regulate a set of genes in a common pathway in order to improve the expression of a desired
protein, and optimize these approaches using a vaccine protein that is commercially made in yeast. In the
second aim, we devise new selections for use in chemostats and turbidostats to identify the effects of small
changes in proteins, in promoters and in untranslated regions of mRNAs. Further, we modify the turbidostat
platform to enable ramped selections, allowing feedback on temperature or drug concentration to optimize
selections. In the third aim, we design new proteins that bind to a protein-protein interface and disrupt an
interaction. These inhibitory proteins will allow us to assess the roles of individual members of large protein
families that are difficult to analyze. Finally, in the fourth aim, we generate new biosensors to detect the levels
of a small molecule and of a protein produced in yeast.
Terms: <1,25-Dihydroxycholecalciferol Receptors><1,25-Dihydroxyvitamin D 3 Receptors><1,25-Dihydroxyvitamin D3 Receptors><9-cis-Retinoic Acid Receptor><Affinity><Anabolism><Assay><Basal Transcription Factor><Basal transcription factor genes><Binding><Binding Proteins><Bioassay><Biologic Assays><Biological Assay><Biology><Biosensor><CFTR><CFTR Protein><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Calciferol-Binding Protein><Calciol><Calcitriol Receptors><Cas nuclease technology><Cell Body><Cells><Chaperone><Cholecalciferol><Cholecalciferol Receptors><Complex><Cystic Fibrosis Transmembrane Conductance Regulator><DNA><DNA Binding Domain><DNA-Binding Protein Motifs><Deoxyribonucleic Acid><Disulfide Interchange Enzyme><Disulfide Isomerase><Drugs><ERp59 PDI><Engineering><Erp59><Family><Feedback><GSBP><Gc Globulin><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genes><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Genome engineering><Glycosylation Site-Binding Protein><Group-Specific Component Globulin><Guide RNA><HPV><HPV L1 protein><HPV Vaccine><Human Papilloma Virus><Human Papilloma Virus Vaccine><Human Papillomavirus><Human papilloma virus HPV L1 protein><Human papilloma virus L1 protein><Human papillomavirus HPV L1 protein><Human papillomavirus L1 protein><Human papillomavirus Vaccine><Hybrids><Individual><Infectious Human Wart Virus><Lead><Libraries><Ligand Binding Protein><Ligand Binding Protein Gene><Ligands><Medication><Messenger RNA><Methodology><Methods><Molecular Chaperones><Molecular Interaction><Mutation><Pathway interactions><Pb element><Pharmaceutic Preparations><Pharmaceutical Preparations><Play><Production><Property><Protein Binding><Protein Binding Domain><Protein Binding Motif><Protein Biosynthesis><Protein Disulfide Isomerase><Protein Engineering><Protein Family><Protein-Protein Interaction Domain><Proteins><RNA Expression><RXR><RXR Protein><Ramp><Receptor Protein><Reporter Genes><Reporting><Retinoic Acid Receptor RXR><Retinoid X Receptors><Ribosomal Peptide Biosynthesis><Ribosomal Protein Biosynthesis><Ribosomal Protein Synthesis><Role><S-S rearrangase><Site><Source><Steroid Compound><Steroids><Sulfhydryl-Disulfide Interchange Enzyme><System><Temperature><Thiol-Disulfide Transhydrogenase><Transcalciferin><Transcription><Transcription Factor Proto-Oncogene><Transcription factor genes><Trypanothione-Glutathione Thioltransferase><Two-Hybrid Assay><Two-Hybrid Method><Two-Hybrid System Technics><Two-Hybrid System Techniques><UTRs><Untranslated Regions><VIT D><VP 16><VP16><Vaccines><Variant><Variation><Viral Gene Products><Viral Gene Proteins><Viral Proteins><VitD3><Vitamin D><Vitamin D 2><Vitamin D 3><Vitamin D 3 Receptors><Vitamin D Receptors><Vitamin D-Binding Globulin><Vitamin D-Binding Protein><Vitamin D2><Vitamin D3><Vitamin D3 Receptor><Vitamins><Yeasts><biological sensor><biosynthesis><bound protein><combinatorial><cystic fibrosis transmembrane regulator><design><designing><drug/agent><dsbA Gene Product><dsbA Protein><dsbC Gene Product><dsbD Gene Product><experiment><experimental research><experimental study><fitness><gRNA><genetic protein engineering><genome mutation><heavy metal Pb><heavy metal lead><improved><inhibitor><inhibitor/antagonist><mRNA><member><novel><pathway><prevent><preventing><promoter><promotor><protein design><protein protein interaction><protein synthesis><receptor><response><small molecule><social role><trafficking><transcription factor><virus protein><wart virus><xprA Gene Product><yeast genome>