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Progressive Loss of Function in a Limb Enhancer during Snake Evolution.

Evgeny Z Kvon1, Olga K Kamneva2, Uirá S Melo1

  • 1MS 84-171, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

Cell
|October 22, 2016
PubMed
Summary
This summary is machine-generated.

Snake evolution involved changes in a limb enhancer controlling Sonic hedgehog (Shh). Restoring a single binding site in this enhancer fully restored limb development, revealing regulatory sequences

Keywords:
CRISPR/Cas9Sonic hedgehog (Shh)ZRScis-regulatory elementenhancerevo-devogenome editinglimb developmentmorphological evolutionsnakes

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Area of Science:

  • Evolutionary developmental biology
  • Comparative genomics
  • Molecular evolution

Background:

  • Body shape evolution is linked to regulatory sequence changes.
  • Molecular mechanisms behind major vertebrate morphological transitions are not well understood.

Purpose of the Study:

  • Investigate molecular events driving major morphological transitions in vertebrates.
  • Identify specific sequence changes in regulatory elements associated with body plan evolution.

Main Methods:

  • Identified snake-specific sequence changes in a conserved Sonic hedgehog (Shh) limb enhancer.
  • Utilized transgenic mouse reporter assays to test enhancer activity in vivo.
  • Performed genomic substitution experiments in mice using orthologous enhancers from different species.
  • Synthetically restored a lost transcription factor binding site.

Main Results:

  • The limb enhancer's activity pattern is conserved in vertebrates like fish but altered in snakes.
  • Replacing mouse enhancer with human or fish orthologs resulted in normal limb development.
  • Snake orthologs of the enhancer caused severe limb reduction in mice.
  • Restoring a single lost transcription factor binding site rescued enhancer function in snakes.

Conclusions:

  • Changes in a specific regulatory sequence are associated with the major body plan transition to snake-like forms.
  • Enhancers play a critical role in morphological evolution.
  • Identified a key regulatory mechanism underlying limb loss in snakes.