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Laboratory Culture and Mutagenesis of Amphioxus (Branchiostoma floridae)

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Book cover Developmental Biology of the Sea Urchin and Other Marine Invertebrates

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2219))

Abstract

Cephalochordates (amphioxus) are invertebrate chordates closely related to vertebrates. As they are evolving very slowly, they are proving to be very appropriate for developmental genetics studies aimed at understanding how vertebrates evolved from their invertebrate ancestors. To date, techniques for gene knockdown and overexpression have been developed, but methods for continuous breeding cultures and generating germline mutants have been developed only recently. Here we describe methods for continuous laboratory breeding cultures of the cephalochordate Branchiostoma floridae and the TALEN and Tol2 methods for mutagenesis. Included are strategies for analyzing the mutants and raising successive generations to obtain homozygotes. These methods should be applicable to any warm water species of cephalochordates with a relatively short generation time of 3–4 months and a life span of 3 years or more.

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Acknowledgments

Particular thanks are due to technicians and many students who helped establish and maintain the amphioxus cultures. These include Ms. Patti Brewer, Ms. Christine Steinke, Mr. Dan Hartsook, Ms. Esmeralda Alcantar, Mr. Joel Lashomb, Ms. Taylor Crabtree, Mr. Zaren Youngblood, Ms. Denisse Larin Enriquez, Mr. Pablo Perez, and a host of volunteers. Special thanks are given to Prof. Yiquan Wang and students Jun Feng and Chenggand Shi for their contributions to the development of genome editing and transgenic methods in amphioxus. Culture of B. floridae was supported by NSF IOS 1160880 to L.Z.H. Development of genome editing and transgenesis of amphioxus was supported by NSFC 31672246 to G.L.

Author information

Authors and Affiliations

  1. Marine Biology Research Division, Scripps Institution of Oceanography, University of California San Diego, La Jolla, CA, USA

    Linda Z. Holland

  2. State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian, China

    Guang Li

Authors
  1. Linda Z. Holland
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  2. Guang Li
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Corresponding authors

Correspondence to Linda Z. Holland or Guang Li .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, Midwestern University, Glendale, AZ, USA

    David J. Carroll

  2. Department of Biology, University of New Mexico, Albuquerque, NM, USA

    Stephen A. Stricker

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Holland, L.Z., Li, G. (2021). Laboratory Culture and Mutagenesis of Amphioxus (Branchiostoma floridae). In: Carroll, D.J., Stricker, S.A. (eds) Developmental Biology of the Sea Urchin and Other Marine Invertebrates. Methods in Molecular Biology, vol 2219. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0974-3_1

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  • DOI: https://doi.org/10.1007/978-1-0716-0974-3_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0973-6

  • Online ISBN: 978-1-0716-0974-3

  • eBook Packages: Springer Protocols

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