Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies

Xueyan Li, Dingding Fan, Wei Zhang, Guichun Liu, Lu Zhang, Li Zhao, Xiaodong Fang, Lei Chen, Yang Dong, Yuan Chen, Yun Ding, Ruoping Zhao, Mingji Feng, Yabing Zhu, Yue Feng, Xuanting Jiang, Deying Zhu, Hui Xiang, Xikan Feng, Shuaicheng Li, Jun Wang, Guojie Zhang, Marcus R. Kronforst () and Wen Wang ()
Additional contact information
Xueyan Li: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Dingding Fan: BGI-Shenzhen
Wei Zhang: University of Chicago
Guichun Liu: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Lu Zhang: BGI-Shenzhen
Li Zhao: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Xiaodong Fang: BGI-Shenzhen
Lei Chen: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Yang Dong: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Yuan Chen: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Yun Ding: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Ruoping Zhao: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Mingji Feng: BGI-Shenzhen
Yabing Zhu: BGI-Shenzhen
Yue Feng: BGI-Shenzhen
Xuanting Jiang: BGI-Shenzhen
Deying Zhu: BGI-Shenzhen
Hui Xiang: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences
Xikan Feng: BGI-Shenzhen
Shuaicheng Li: City University of Hongkong
Jun Wang: BGI-Shenzhen
Guojie Zhang: BGI-Shenzhen
Marcus R. Kronforst: University of Chicago
Wen Wang: State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences

Nature Communications, 2015, vol. 6, issue 1, 1-10

Abstract: 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.

Date: 2015
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DOI: 10.1038/ncomms9212

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