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Multiplex clonal analysis in the chick embryo using retrovirally-mediated combinatorial labeling

Tang, Weiyi and Li, Yuwei and Gandhi, Shashank and Bronner, Marianne E. (2019) Multiplex clonal analysis in the chick embryo using retrovirally-mediated combinatorial labeling. Developmental Biology, 450 (1). pp. 1-8. ISSN 0012-1606. http://resolver.caltech.edu/CaltechAUTHORS:20190315-101353338

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Abstract

Lineage analysis plays a central role in exploring the developmental potential of stem and progenitor cell populations. In higher vertebrates, a variety of techniques have been used to label individual cells or cell populations, including interspecies grafting, intracellular microinjection, and Cre-mediated recombination. However, these approaches often suffer from difficulties in progenitor cell targeting, low cellular resolution and/or ectopic labeling. To circumvent these issues, here we utilize replication incompetent avian (RIA) retroviruses to deliver combinations of fluorescent proteins into distinct cellular compartments in chick embryos. In particular, RIA-mediated lineage tracing is optimal for long term mapping of dispersing cell populations like the neural crest. Using this tool, we confirm that trunk neural crest cells are multipotent. Furthermore, our RIA vector is engineered to be fully adaptable for other purposes such as cell fate analysis, gene perturbation studies and time-lapse imaging. Taken together, we present a novel approach of multiplex lineage analysis that can be applied to normal and perturbed development of diverse cell populations in avian embryos.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.ydbio.2019.03.007DOIArticle
ORCID:
AuthorORCID
Li, Yuwei0000-0001-7753-4869
Gandhi, Shashank0000-0002-4081-4338
Bronner, Marianne E.0000-0003-4274-1862
Additional Information:© 2019 Published by Elsevier Inc. Received 6 December 2018, Revised 11 March 2019, Accepted 12 March 2019, Available online 15 March 2019. Author contributions: W.T., Y.L. and M.E.B conceived the project. W.T., Y.L. and S.G performed the experiments. W.T., Y.L. and M.E.B wrote the paper with consultation from S.G. This work is supported by NIHR01DE027568 and NIHRO1HL14058 to M.E.B and American Heart Association predoctoral fellowship 18PRE34050063 to S.G. The authors declare no competing interests. We thank Alison Koontz for providing insightful suggestions on probability calculation, Can Li, Felipe Vieceli, David Miller for technical support. We thank Carlos Lois laboratory and Caltech Biological Imaging Facility for sharing equipment.
Funders:
Funding AgencyGrant Number
NIHR01DE027568
NIHR01HL14058
American Heart Association18PRE34050063
Record Number:CaltechAUTHORS:20190315-101353338
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20190315-101353338
Official Citation:Weiyi Tang, Yuwei Li, Shashank Gandhi, Marianne E. Bronner, Multiplex clonal analysis in the chick embryo using retrovirally-mediated combinatorial labeling, Developmental Biology, Volume 450, Issue 1, 2019, Pages 1-8, ISSN 0012-1606, https://doi.org/10.1016/j.ydbio.2019.03.007. (http://www.sciencedirect.com/science/article/pii/S0012160618307802)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:93862
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:15 Mar 2019 17:19
Last Modified:24 Apr 2019 16:57

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