Source: WEST VIRGINIA STATE UNIVERSITY submitted to
SPATIAL TRANSCRIPTOMIC SEQUENCING AND TRIP-CRISPR KNOCKOUT TECHNOLOGIES FOR TEACHING AND EXPLORING THE HEALTH EFFECTS OF CAPSAICIN, TURMERIC, BLUEBERRIES, AND BITTER MELON USING DROSOPHILA
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
Reporting Frequency
Annual
Accession No.
1031945
Grant No.
2024-38821-42101
Project No.
wvax-SpacTrans24
Proposal No.
2023-09267
Multistate No.
(N/A)
Program Code
EQ
Project Start Date
Apr 1, 2024
Project End Date
Mar 31, 2027
Grant Year
2024
Project Director
Hankins, G.
Recipient Organization
WEST VIRGINIA STATE UNIVERSITY
PO BOX 1000
INSTITUTE,WV 25112
Performing Department
(N/A)
Non Technical Summary
This project explores the fascinating world of phytochemicals--naturally occurring compounds in fruits, vegetables, and spices known for their health benefits and low toxicity. Phytochemicals such as polyphenols, carotenoids, flavonoids, curcuminoids, terpenoids, and capsaicinoids are not only integral to our diet but also play a crucial role in the growth and health of organisms. To unravel how these compounds affect health, we turn to the fruit fly, Drosophila melanogaster, a tiny but powerful model organism. With a genetic makeup surprisingly similar to humans, fruit flies offer a window into understanding how our bodies might react to different diets. In this project, we employ cutting-edge science, including Spatial Transcriptomic Sequencing and CRISPR gene-editing technology, to delve into the effects of consuming capsaicin, turmeric, blueberries, and bittermelon. Our research begins with detailed investigation into these phytochemicals, followed by designing and conducting experiments that expose fruit flies to these compounds. We then use advanced sequencing techniques to study changes in gene expression and CRISPR technology to investigate the roles of specific genes. Additionally, we're harnessing the power of Artificial Intelligence (AI) to revolutionize how we observe and analyze the fruit flies' behaviors after consuming these compounds. By automating data collection and employing AI for image recognition and pattern analysis, we aim to gain deeper insights with less manual effort. This innovative approach promises to shed new light on the molecular interactions and health implications of consuming natural phytochemicals, offering a glimpse into the future of dietary research and health science.
Animal Health Component
0%
Research Effort Categories
Basic
50%
Applied
30%
Developmental
20%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
30250101080100%
Knowledge Area
302 - Nutrient Utilization in Animals;

Subject Of Investigation
5010 - Food;

Field Of Science
1080 - Genetics;
Goals / Objectives
The objectives of this research project are to employ spatial transcriptomics for the investigation of genome-wide responses in Drosophila melanogaster (fruit flies) subjected to diets enriched with phytochemicals. This will entail a comprehensive analysis of gene expression patterns across different tissues to elucidate the molecular mechanisms underlying the dietary impact of phytochemicals on organismal health and development.Further, the project aims to generate mutant D. melanogaster lines for genes identified as key responders to phytochemical exposure. This will be achieved using the Targeted RNAi Interference Project (TRiP)-CRISPR/Cas9 system to specifically knock out causal genes, enabling a detailed functional analysis of these genes in the context of phytochemical intake.Incorporating machine learning techniques, the project will analyze physiological and behavioral responses of these genetically modified flies to different phytochemical diets. This approach will leverage advanced computational models to discern patterns and predict outcomes from complex datasets, including phenotypic observations and high-throughput genetic data.Lastly, the project proposes the integration of these research themes into a structured educational framework for a Summer School program targeted at undergraduate and high school students. This will involve hands-on research projects that cover aspects of behavior, physiology, transcriptomics, and genetic validation techniques. The aim is to provide a comprehensive educational experience that bridges theoretical knowledge with practical research skills, fostering the next generation of scientists in the field of genetic and molecular biology.
Project Methods
The plan of operation and methodology detailed for studying the genome-wide responses in Drosophila melanogaster fed with phytochemicals encompasses several sophisticated techniques. Initially, adult Drosophila melanogaster are fed a diet enriched with specific phytochemicals, such as capsaicin, antioxidants, and curcumin, over a set period. Following this, tissues like the brain, gut, and muscles are isolated, fixed, and prepared for spatial transcriptomics analysis, which includes histological staining, RNA isolation, library preparation, sequencing, and data analysis to identify differentially expressed genes. This comprehensive approach integrates cutting-edge molecular biology techniques, including next-generation sequencing and bioinformatics tools, to map and analyze gene expression data spatially, aiming to uncover the genetic responses to phytochemical exposure.In addition to transcriptomic analysis, the project outlines developing Drosophila mutants via Trip-CRISPR knockout and utilizing machine learning to study the physiology and behavior of these mutants under phytochemical diets. Behavioral assays, metabolic measurements, and genomic data collection are followed by data cleaning, feature engineering, and the application of machine learning models to interpret the physiological and behavioral impacts of dietary phytochemicals on Drosophila mutants. The methodology extends to a broader educational component, where independent research projects related to Drosophila's behavior, physiology, transcriptomics, and mutant validation are proposed for a Summer School for undergraduate and high school students. This multi-faceted approach aims to foster multidisciplinary collaborations and generate insights into the effects of phytochemicals on health, emphasizing the project's ambition to bridge human nutrition, genomics, metabolomics, biomedical research, and artificial intelligence to advance obesity research and identify health-enhancing phytochemicals.