Presentation/Session Information

Session Information

Session Title: RNA Interference, Noncoding RNAs, and Genetic Technologies Session Type: Parallel
Session Location: Carnesale Palisades Ballroom Session Time: Sat, Jun 27 8:30AM - 11:30AM

Presentation Information

Program Number: 148 Presentation Time: 10:54AM - 11:06AM

Presentation Content

Computer-assisted transgenesis of C. elegans for deep phenotyping.Cody Gilleland 1, Adam Falls 1, James Noraky 1, Maxwell Heiman 2, Mehmet Yanik 1,3,4. 1)Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA; 2)Department of Genetics, Boston Children<sym07>s Hospital and Harvard Medical School, Boston, MA; 3)Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA; 4)Department of Electrical Engineering, ETH Zürich, 8093 Zürich, Switzerland

A major goal in the study of human disease is to assign functions to genes or genetic variants. The model organism C. elegans provides a powerful tool because homologs of many human genes are identifiable, and large collections of genetic vectors and mutant strains are available.  However, the delivery of such vector libraries into mutant strains remains a long-standing experimental bottleneck for phenotypic analysis. Here, we present a computer-assisted micro-injection (CAMI) platform to streamline the production of transgenic C. elegans with multiple vectors for deep phenotyping. Briefly, animals are immobilized in a temperature-sensitive hydrogel using a standard multiwell platform. Microinjection is then performed under control of an automated microscope using precision robotics driven by customized computer vision algorithms. We demonstrate utility by phenotyping the morphology of 12 neuronal classes in 6 mutant backgrounds using combinations of neuron-type-specific fluorescent reporters.  A standard assay of rol-6 plasmid expression efficiency determined that transgenesis by CAMI occurs at an efficiency well in line with conventional techniques. CAMI operates at a rate of ~25 sec per gonad target with no user fatigue, in contrast to conventional manual microinjection at ~2-3 min per animal where, in addition, most users cannot sustain injecting for more than 1-2 hours. Typically, one needs to inject ~15 animals to ensure obtaining a transgenic line. Importantly, however, recent genome-editing techniques such as Crispr/Cas9-mediated genome editing require larger cohorts of ~50 animals, which are challenging using manual methods but are easily accommodated using CAMI. This technology can industrialize the assignment of in vivo gene function by enabling large-scale transgenic engineering.




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