Researchers at New York University have developed a new protocol to generate gene-specific GAL4 lines in Drosophila melanogaster (fruit fly) through genetic crosses, offering a cost-effective alternative to traditional microinjection methods. The team, led by Dr. Yu-Chieh David Chen, and including Siqi April Li, Hongzhou Gustave Li, Nathalie Shoji, and Dr. Claude Desplan, detailed their innovative approach in the journal STAR Protocols.
Dr. Chen said, “This protocol significantly reduces the cost and complexity of creating gene-specific GAL4 lines, making it accessible to fly researchers focusing on multiple tissues.” The work was published in the peer-reviewed journal, STAR Protocols.
The researchers described a step-by-step method that includes four rounds of in vivo genetic crosses, PCR genotyping, and fluorescence imaging to ensure correct targeting of GAL4 integration before establishing stable fly populations. This process avoids the need for microinjection, making it more accessible and less expensive.
One of the key findings of the study is that this new method is adaptable and can be used to generate a large batch of split GAL4 hemi-conductors, which is essential for researchers working on different tissues in Drosophila. The researchers demonstrated the efficacy of their protocol by successfully generating split GAL4 lines from existing MiMIC/CRIMIC lines, which are widely used in fruit fly genetic studies.
Professor Desplan highlighted the importance of this breakthrough: “Our approach offers a significant advantage over traditional methods, as it reduces the time and resources required to create gene-specific drivers. This will accelerate research in several fields, including neurobiology, developmental biology and genetics.”
In their study, the researchers also highlighted the importance of phenotypic markers for tracking transgenes during genetic crosses. They noted that incorrect markers could indicate contamination of populations, which could compromise results. The protocol includes detailed instructions for maintaining fly populations and preparing the necessary reagents and buffer solutions.
Furthermore, the entire protocol can be employed as part of Course-Based Undergraduate Research Experiences (CUREs), which provide hands-on experience in conducting original fly genetics research. Lines generated from the protocol will be broadly useful for generating cell-type-specific split GAL4 lines, to the benefit of researchers in the fly community.
In conclusion, the new protocol developed by Dr. Chen and colleagues represents a significant advance in the field of Drosophila genetics. By offering a cost-effective and efficient alternative to traditional methods, this protocol is set to enhance the capabilities of researchers worldwide, enabling more extensive and accurate genetic studies.
Journal reference
Li, S.A., Li, H.G., Shoji, N., Desplan, C., and Chen, Y.D. “Protocol for replacing MiMIC and CRIMIC intronic coding lines with T2A-split-GAL4 lines in Drosophila by genetic crosses”. STAR Protocols, 2023. DOI: https://doi.org/10.1016/j.xpro.2023.102706
About the authors
David Chen, Yu-Chieh's sonDavid is an NIH F32 and K99/R00 Fellow in the laboratory of Dr. Claude Desplan at New York University. David was a Howard Hughes Medical Institute (HHMI) International Research Scholar and did his graduate studies in the laboratory of Dr. Anupama Dahanukar at the University of California Riverside. His long-term research interest is in understanding the developmental processes and functional roles of the primary senses: vision, touch, hearing, and chemosensitivity (taste and smell), which are essential for organisms to perceive and interact with the world. David is currently on the faculty job market, looking to find a place to launch his independent research program. His future research program lies in understanding the molecular regulators underlying various developmental events, such as cell specification, axonal guidance, and synaptic partner matching, to form the functional sensory circuits in DrosophilaDavid is also passionate about teaching and mentoring the next generation of scientists. Please visit his website for more information: ycdavidchen.com
Siqi (April) LiSiqi earned her BA in Biology from New York University's College of Arts and Sciences. She is particularly passionate about molecular biology, as she seeks to understand how microscopic physical processes govern the behavior of biological systems at all possible scales. In the past, Siqi interned at a pharmaceutical company, where she was tasked with creating an online platform that educates the general public about discoveries found in the academic literature. More recently, Siqi has worked in a neuropsychology lab as a research volunteer, assisting with a wide range of experimental procedures. She joined the Desplan lab in the fall of 2022, working with David Chen to generate split GAL4 genetic tools to label new cell types in development. Drosophila Visual system. Inspired by the people she met along this journey, Siqi is pursuing her PhD at the Icahn School of Medicine at Mount Sinai starting in the fall of 2024.
Hongzhou (Gustave) Li Gustave earned a BSc in Neuroscience with honors from NYU Shanghai in 2024. He became interested in neurobiology from the intricate biological pathways that contribute to brain development and function, and is passionate about understanding the brain from a bottom-up perspective. During his undergraduate studies, Gustave worked in the lab of Dr. Claude Desplan at NYU Biology and was directly mentored by postdoctoral researcher Dr. Yu-Chieh David Chen, where he conducted research on generating genetic tools that label individual neuronal cell types in the Drosophila central nervous system. He presented his work at multiple academic conferences and was awarded the Best Poster Presentation in Developmental Biology and Genetics at the 2023 Annual Biomedical Research Conference for Minority Scientists (ABRCMS). Beginning in Fall 2024, he will pursue a PhD in Neurobiology at the California Institute of Technology. He plans to enter academia and become an independent researcher after his PhD.
You have successfully subscribed!
