Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies.
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.
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Xueyan
http://orcid.org/0000-0003-0457-7846
X
Li
BGI-Shenzhen, Shenzhen 518083, China.
Dingding
D
Fan
Department of Ecology and Evolution, University of Chicago, Chicago, Illinois 60637, USA.
Wei
W
Zhang
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Guichun
G
Liu
BGI-Shenzhen, Shenzhen 518083, China.|City University of Hongkong, Hongkong, China.
Lu
L
Zhang
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.|Department of Evolution and Ecology, University of California Davis, Davis, California 95616, USA.
Li
L
Zhao
BGI-Shenzhen, Shenzhen 518083, China.
Xiaodong
X
Fang
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.|University of Chinese Academy of Sciences, Beijing 100049, China.
Lei
L
Chen
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Yang
Y
Dong
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Yuan
Y
Chen
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Yun
Y
Ding
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Ruoping
R
Zhao
BGI-Shenzhen, Shenzhen 518083, China.
Mingji
M
Feng
BGI-Shenzhen, Shenzhen 518083, China.
Yabing
Y
Zhu
BGI-Shenzhen, Shenzhen 518083, China.
Yue
Y
Feng
BGI-Shenzhen, Shenzhen 518083, China.
Xuanting
X
Jiang
BGI-Shenzhen, Shenzhen 518083, China.|School of Bioscience and Biotechnology, South China University of Technology, Guangzhou 510641, China.
Deying
D
Zhu
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Hui
H
Xiang
BGI-Shenzhen, Shenzhen 518083, China.|City University of Hongkong, Hongkong, China.
Xikan
X
Feng
City University of Hongkong, Hongkong, China.
Shuaicheng
S
Li
BGI-Shenzhen, Shenzhen 518083, China.
Jun
J
Wang
BGI-Shenzhen, Shenzhen 518083, China.|Centre for Social Evolution, Department of Biology, Universitetsparken 15, University of Copenhagen, Copenhagen DK-2100, Denmark.
Guojie
G
Zhang
Department of Ecology and Evolution, University of Chicago, Chicago, Illinois 60637, USA.
Marcus R
MR
Kronforst
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.
Wen
W
Wang
D000818:Animals; D001483:Base Sequence; D002080:Butterflies / Q000235:genetics*; D064113:CRISPR-Cas Systems; D004268:DNA-Binding Proteins / Q000235:genetics*; D051157:Frizzled Receptors / Q000235:genetics*; D020869:Gene Expression Profiling; D005821:Genetic Techniques; D014644:Genetic Variation; D049750:Genome, Insect / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D019476:Insect Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010641:Phenotype
D004268:DNA-Binding Proteins; D051157:Frizzled Receptors; D018398:Homeodomain Proteins; D019476:Insect Proteins