RNA interference can be used to disrupt gene function in tardigrades

How morphological diversity arises is a key question in evolutionary developmental biology. As a long-term approach to address this question, we are developing the water bear Hypsibius dujardini (Phylum Tardigrada) as a model system. We expect that using a close relative of two well-studied models,...

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Bibliographic Details
Published in:Development Genes and Evolution Vol. 223; no. 3; pp. 171 - 181
Main Authors: Tenlen, Jennifer R., McCaskill, Shaina, Goldstein, Bob
Format: Journal Article
Language:English
Published: Berlin/Heidelberg Springer-Verlag 01.05.2013
Springer Nature B.V
Subjects:
Animal Genetics and Genomics
Animals
Arthropoda
Biomedical and Life Sciences
Caenorhabditis elegans
Cell Biology
classification
Cloning, Molecular
Developmental Biology
Double-stranded RNA
Drosophila
Embryogenesis
evolution
Evolutionary Biology
Evolutionary genetics
gene targeting
Genes
Genes, Lethal
genetics
Green fluorescent protein
Hypsibius dujardini
Lethality
Life Sciences
messenger RNA
mRNA
Nematoda
Original Article
phenotype
Phenotypes
Phylogeny
Plant Genetics and Genomics
reverse transcriptase polymerase chain reaction
RNA Interference
RNA-mediated interference
Tardigrada
Tardigrada - classification
Tardigrada - genetics
Zoology
Evo-devo
RNA interference
Tardigrade
Extreme environments
ISSN:0949-944X, 3059-3239, 1432-041X, 1432-041X, 3059-3247
Online Access:Get full text
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Summary:How morphological diversity arises is a key question in evolutionary developmental biology. As a long-term approach to address this question, we are developing the water bear Hypsibius dujardini (Phylum Tardigrada) as a model system. We expect that using a close relative of two well-studied models, Drosophila (Phylum Arthropoda) and Caenorhabditis elegans (Phylum Nematoda), will facilitate identifying genetic pathways relevant to understanding the evolution of development. Tardigrades are also valuable research subjects for investigating how organisms and biological materials can survive extreme conditions. Methods to disrupt gene activity are essential to each of these efforts, but no such method yet exists for the Phylum Tardigrada. We developed a protocol to disrupt tardigrade gene functions by double-stranded RNA-mediated RNA interference (RNAi). We showed that targeting tardigrade homologs of essential developmental genes by RNAi produced embryonic lethality, whereas targeting green fluorescent protein did not. Disruption of gene functions appears to be relatively specific by two criteria: targeting distinct genes resulted in distinct phenotypes that were consistent with predicted gene functions and by RT-PCR, RNAi reduced the level of a target mRNA and not a control mRNA. These studies represent the first evidence that gene functions can be disrupted by RNAi in the phylum Tardigrada. Our results form a platform for dissecting tardigrade gene functions for understanding the evolution of developmental mechanisms and survival in extreme environments.
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Present address: Dept. of Biology, Seattle Pacific University, 3307 3rd Ave W., Suite 205, Seattle, WA 98119, USA
ISSN:0949-944X
3059-3239
1432-041X
1432-041X
3059-3247
DOI:10.1007/s00427-012-0432-6