Article open access publication

Outbred genome sequencing and CRISPR/Cas9 gene editing in butterflies

Nature Communications, Springer Nature, ISSN 2041-1723

Volume 6, 1, 2015

DOI:10.1038/ncomms9212, Dimensions: pub.1049702511, PMC: PMC4568561, PMID: 26354079,


Zhang, Lei (2) (4)
Zhao, Li (1) (5)
Chen, Lei (1) (6)
Dong, Yang (1) (7)
Chen, Yuan (1) (8)
Ding, Yun (1) (9)
Feng, Yue (2)
Zhu, Deying (2) (10)
Feng, Xikan (2) (4)
Wang, Jun (2)
Zhang, Guo-Jie (2) (11)
Wang, Wen * (1)

* Corresponding author



  1. (1) Kunming Institute of Zoology, grid.419010.d
  2. (2) Beijing Genomics Institute, grid.21155.32
  3. (3) University of Chicago, grid.170205.1
  4. (4) University of Hong Kong, grid.194645.b
  5. (5) University of California, Davis, grid.27860.3b
  6. (6) University of Chinese Academy of Sciences, grid.410726.6
  7. (7) Kunming University of Science and Technology, grid.218292.2
  8. (8) Duke University Hospital, grid.189509.c
  9. (9) Howard Hughes Medical Institute, grid.413575.1
  10. (10) South China University of Technology, grid.79703.3a
  11. (11) University of Copenhagen, grid.5254.6, KU


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.


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