Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping

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. Howeve...

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Main Authors:	Gilleland, Cody L., Falls, Adam T., Noraky, James, Heiman, Maxwell G., Yanik, Mehmet F.
Format:	Online
Language:	English
Published:	Genetics Society of America 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4566274/

id	pubmed-4566274
recordtype	oai_dc
spelling	pubmed-45662742015-09-14 Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping Gilleland, Cody L. Falls, Adam T. Noraky, James Heiman, Maxwell G. Yanik, Mehmet F. Investigations 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. Genetics Society of America 2015-09 2015-07-10 /pmc/articles/PMC4566274/ /pubmed/26163188 http://dx.doi.org/10.1534/genetics.115.179648 Text en Copyright © 2015 by the Genetics Society of America Available freely online through the author-supported open access option.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Gilleland, Cody L. Falls, Adam T. Noraky, James Heiman, Maxwell G. Yanik, Mehmet F.
spellingShingle	Gilleland, Cody L. Falls, Adam T. Noraky, James Heiman, Maxwell G. Yanik, Mehmet F. Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
author_facet	Gilleland, Cody L. Falls, Adam T. Noraky, James Heiman, Maxwell G. Yanik, Mehmet F.
author_sort	Gilleland, Cody L.
title	Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
title_short	Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
title_full	Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
title_fullStr	Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
title_full_unstemmed	Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping
title_sort	computer-assisted transgenesis of caenorhabditis elegans for deep phenotyping
description	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.
publisher	Genetics Society of America
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4566274/
_version_	1613474764536414208

Computer-Assisted Transgenesis of Caenorhabditis elegans for Deep Phenotyping

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