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Video Q&A ViewAttendees 6

Physiology, Biochemistry, and Toxicology

10-Minute Paper

PBT: Genetics On-Demand Presentations

CRISPR/Cas9 and chromatin accessibility reveal regulatory switches of WntA: A butterfly wing pattern ground-plan gene

On-Demand
On-Demand

    Presenting Author(s)

  • Anyi Mazo-Vargas

    Ford Foundation Postdoctoral Fellow
    The George Washington University
    Washington, District of Columbia

    • Co-Author(s)

  • Arnaud Martin

    Assistant Professor
    The George Washington University
    Washington, District of Columbia

  • RR

    Robert Reed

    Principal Investigator
    Cornell University
    Ithaca, New York

Butterfly wing patterns are a collection of stripes and shapes that are all features of a conserved groundplan system. These patterns, which are highly diverse and rapidly evolving, are determined through the activity of key developmental patterning molecules that delimit specific groundplan elements. In particular, the gene WntA plays a crucial role in marking color pattern boundaries in both nymphalids and papilionids. How can a single gene establish the diversity of color patterns we see in these insects? One hypothesis is that gene regulatory regions can encode deep pattern homology while also driving adaptive change. Here, we characterized the regulatory architecture of the WntA gene in nymphalid butterflies. First, we defined chromatin accessibility profiles across five species of nymphalids to identify candidate cis-regulatory elements (CREs). Then, we used CRISPR/Cas9 in vivo knockouts of 46 CREs to test their functions in wing patterning. Our results revealed extensive homology of groundplan CREs across nymphalids, except for in monarch butterflies. Independent deletions of individual CREs consistently resulted in strong, widespread, and overlapping color pattern effects, and most CREs showed both enhancer- and silencer-like functionality. Our results suggest that color pattern evolution likely occurred through changes to a collection of deeply conserved, interacting groundplan CREs, as opposed to rapid turnover and divergence of pattern-specific CREs. Moreover, our findings highlight the multifunctionality of pattern formation CREs, and how they appear to work in highly interdependent networks.