title: Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping.
abstract: A major goal in the study of human diseases is to assign functions to genes or genetic variants. The model organism Caenorhabditis 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 microinjection 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. Microinjections are 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 six mutant backgrounds using combinations of neuron-type-specific fluorescent reporters. This technology can industrialize the assignment of in vivo gene function by enabling large-scale transgenic engineering.
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
authors.forename: Cody L
authors.initials: CL
authors.lastname: Gilleland
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
authors.forename: Adam T
authors.initials: AT
authors.lastname: Falls
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139.
authors.forename: James
authors.initials: J
authors.lastname: Noraky
authors.affiliation: Department of Genetics, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115 heiman@genetics.med.harvard.edu yanikm@ethz.ch.
authors.forename: Maxwell G
authors.identifier: http://orcid.org/0000-0002-2557-6490
authors.initials: MG
authors.lastname: Heiman
authors.affiliation: Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 Department of Information Technology and Electrical Engineering, ETH, 8057 Zürich, Switzerland heiman@genetics.med.harvard.edu yanikm@ethz.ch.
authors.forename: Mehmet F
authors.identifier: http://orcid.org/0000-0002-8963-2893
authors.initials: MF
authors.lastname: Yanik
mesh_terms: D000465:Algorithms; D000818:Animals; D030801:Animals, Genetically Modified; D057205:Automation, Laboratory; D017173:Caenorhabditis elegans / Q000235:genetics*; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D006801:Humans; D008845:Microinjections / Q000295:instrumentation* / Q000379:methods; D010641:Phenotype; D012371:Robotics / Q000379:methods*
keywords: C. elegans; automation; deep phenotyping; high throughput; transgenesis