Lineage tracing of genome-edited alleles reveals high fidelity axolotl limb regeneration

  1. Grant Parker Flowers  Is a corresponding author
  2. Lucas D Sanor
  3. Craig M Crews  Is a corresponding author
  1. Yale University, United States

Abstract

Salamanders are unparalleled among tetrapods in their ability to regenerate many structures, including entire limbs, and the study of this ability may provide insights into human regenerative therapies. The complex structure of the limb poses challenges to the investigation of the cellular and molecular basis of its regeneration. Using CRISPR/Cas, we genetically labelled unique cell lineages within the developing axolotl embryo and tracked the frequency of each lineage within amputated and fully regenerated limbs. This allowed us, for the first time, to assess the contributions of multiple low frequency cell lineages to the regenerating limb at once. Our comparisons reveal that regenerated limbs are high fidelity replicas of the originals even after repeated amputations.

Article and author information

Author details

  1. Grant Parker Flowers

    Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, United States
    For correspondence
    grant.flowers@yale.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7436-3531
  2. Lucas D Sanor

    Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Craig M Crews

    Department of Molecular, Cell and Developmental Biology, Yale University, New Haven, United States
    For correspondence
    craig.crews@yale.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8456-2005

Funding

Connecticut Innovations (Seed Grant 15RMA-YALE-09)

  • Grant Parker Flowers

Eunice Kennedy Shriver National Institute of Child Health and Human Development (Individual Postdoctoral Fellowship F32HD086942)

  • Grant Parker Flowers

Connecticut Innovations (Established Investigator Award 15-RMB-YALE-01)

  • Craig M Crews

National Institute of General Medical Sciences (Predoctoral Training Fellowship T32GM007499)

  • Lucas D Sanor

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Experimental procedures were approved by the Yale University IACUC (2016-10557) and were in accordance with all federal policies and guidelines governing the use of vertebrate animals.

Reviewing Editor

  1. Alejandro Sánchez Alvarado, Stowers Institute for Medical Research, United States

Publication history

  1. Received: February 3, 2017
  2. Accepted: September 8, 2017
  3. Accepted Manuscript published: September 16, 2017 (version 1)
  4. Version of Record published: September 29, 2017 (version 2)
  5. Version of Record updated: November 14, 2017 (version 3)

Copyright

© 2017, Flowers et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Grant Parker Flowers
  2. Lucas D Sanor
  3. Craig M Crews
(2017)
Lineage tracing of genome-edited alleles reveals high fidelity axolotl limb regeneration
eLife 6:e25726.
https://doi.org/10.7554/eLife.25726

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