During animal growth and development, fertilized egg cells must divide and give rise to distinct types of cells (e.g. muscle, skin, neuron). Errors in this process result in developmental defects and disease in humans. In order to understand how an animal’s developmental program is encoded by its DNA, it is useful introduce changes in the DNA and analyze the effects on development. At present, however, it is only possible to generate and analyze a few genetic changes at a time in multicellular animals. To overcome this limitation, researchers at North Carolina State University (NCSU) will rapidly probe how cells adopt their fates by quick evolution of small DNA segments that help control gene expression in fly embryos, a good model for other animals. In parallel, the researchers will develop a device for high-throughput analysis of fly embryo development. This project will be undertaken by graduate students at NCSU as part of their research training, and the outcomes will be communicated through workshops and journal articles.

The rapid evolution of small, targeted stretches of DNA will be achieved through repeated rounds of transcription, reverse transcription, and reintegration in live Drosophila cells and embryos. The error-prone process of reverse transcription will introduce mutations into a DNA sequence that is designed to drive the expression of a GFP-tagged reporter gene. To screen a large number of Drosophila embryos for mutations that affect gene expression, a high-throughput platform consisting of a microfluidic array and automated imaging protocols will be constructed. Live embryos will be imaged, and machine learning algorithms will detect and sort embryos that have altered gene expression patterns. The sorted embryos will be harvested for further study. Beyond this study, the development of the high-throughput platform will enable screening protocols in Drosophila embryos, and also in other animal species. This technology can open new avenues in biological design and engineering.

This project is jointly funded by the Division of Molecular and Cellular Biosciences - Genetic Mechanisms and Systems and Synthetic Biology Clusters.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

National Science Foundation (NSF)
Division of Molecular and Cellular Biosciences (MCB)
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Stephen DiFazio
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North Carolina State University Raleigh
United States
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