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Single Cell Transfection in Chick Embryos
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Multiplex clonal analysis in the chick embryo using retrovirally-mediated combinatorial labeling.

Weiyi Tang1, Yuwei Li1, Shashank Gandhi1

  • 1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.

Developmental Biology
|March 20, 2019
PubMed
Summary

Replication incompetent avian retroviruses enable precise lineage tracing in chick embryos. This method confirms trunk neural crest cells are multipotent and offers adaptability for diverse developmental studies.

Keywords:
Combinatorial labellingMultiplexed clonal analysisMultipotencyNeural tubeReplication incompetent avian retrovirusTrunk neural crest

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Area of Science:

  • Developmental Biology
  • Genetics
  • Cell Biology

Background:

  • Lineage analysis is crucial for understanding stem and progenitor cell development.
  • Existing methods like grafting and microinjection have limitations in targeting, resolution, and specificity.
  • Neural crest cell development is a key area of study in vertebrate embryogenesis.

Purpose of the Study:

  • To develop a novel, high-resolution lineage tracing technique for avian embryos.
  • To overcome limitations of existing lineage analysis methods, such as progenitor cell targeting and ectopic labeling.
  • To investigate the multipotency of trunk neural crest cells in chick embryos.

Main Methods:

  • Utilized replication incompetent avian (RIA) retroviruses for delivering fluorescent proteins into chick embryo cells.
  • Employed RIA-mediated lineage tracing for long-term mapping of dispersing cell populations.
  • Applied the technique to study trunk neural crest cell populations.

Main Results:

  • Demonstrated the efficacy of RIA retroviruses for precise, long-term lineage tracing in avian embryos.
  • Confirmed the multipotency of trunk neural crest cells using the developed RIA method.
  • Showcased the adaptability of the RIA vector for various applications including cell fate analysis and gene perturbation.

Conclusions:

  • RIA retroviruses provide a powerful and versatile tool for multiplex lineage analysis in avian development.
  • This novel approach enhances the study of normal and perturbed development in diverse cell populations.
  • The technique offers significant advantages over traditional methods for studying complex developmental processes.