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Nat Commun. 2015 Sep 10;6:8212. doi: 10.1038/ncomms9212.

Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies.

Author information

1
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
2
BGI-Shenzhen, Shenzhen 518083, China.
3
Department of Ecology and Evolution, University of Chicago, Chicago, Illinois 60637, USA.
4
City University of Hongkong, Hongkong, China.
5
Department of Evolution and Ecology, University of California Davis, Davis, California 95616, USA.
6
University of Chinese Academy of Sciences, Beijing 100049, China.
7
School of Bioscience and Biotechnology, South China University of Technology, Guangzhou 510641, China.
8
Centre for Social Evolution, Department of Biology, Universitetsparken 15, University of Copenhagen, Copenhagen DK-2100, Denmark.

Abstract

Butterflies are exceptionally diverse but their potential as an experimental system has been limited by the difficulty of deciphering heterozygous genomes and a lack of genetic manipulation technology. Here we use a hybrid assembly approach to construct high-quality reference genomes for Papilio xuthus (contig and scaffold N50: 492 kb, 3.4 Mb) and Papilio machaon (contig and scaffold N50: 81 kb, 1.15 Mb), highly heterozygous species that differ in host plant affiliations, and adult and larval colour patterns. Integrating comparative genomics and analyses of gene expression yields multiple insights into butterfly evolution, including potential roles of specific genes in recent diversification. To functionally test gene function, we develop an efficient (up to 92.5%) CRISPR/Cas9 gene editing method that yields obvious phenotypes with three genes, Abdominal-B, ebony and frizzled. Our results provide valuable genomic and technological resources for butterflies and unlock their potential as a genetic model system.

PMID:
26354079
PMCID:
PMC4568561
DOI:
10.1038/ncomms9212
[Indexed for MEDLINE]
Free PMC Article

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