Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping.

Genetics (2015-07-15)
Cody L Gilleland, Adam T Falls, James Noraky, Maxwell G Heiman, Mehmet F Yanik
RESUMEN

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.

MATERIALES
Referencia del producto
Marca
Descripción del producto

Sigma-Aldrich
DIC, 99%
Sigma-Aldrich
Diisopropylcarbodiimide solution, 1 M in dichloromethane
Sigma-Aldrich
Diisopropylcarbodiimide solution, 1 M in THF