Microinjection of Exogenous Nucleic Acids into Eggs: Ciona Species

  • Kenji KobayashiEmail author
  • Yutaka Satou
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1029)


Microinjection is a common technique used to deliver nucleic acids into eggs and embryos in Ciona species. There are three Ciona species that are commonly used for research—Ciona intestinalis type A (C. robusta), C. intestinalis type B (C. intestinalis), and C. savignyi. Here, we present the microinjection methods using eggs and embryos of C. intestinalis type A and C. savignyi; however, our methods would also be applicable to eggs and embryos of C. intestinalis type B. Microinjection is a classical and widely used delivery method, which involves the use of a glass micropipette, a hollow glass needle with a microscopic tip, to inject nucleic acids into eggs and embryos under a stereo microscope. The required amount of nucleic acids is much smaller for microinjection than for electroporation, another delivery method. Proteins, and other chemicals, such as fluorescent dye, can be introduced with nucleic acids using a microinjection.


Developmental biology Microinjection Ascidian Ciona Nucleic acid DNA RNA Morpholino oligonucleotide Egg Embryo 


  1. Brunetti R, Gissi C, Pennati R, Caicci F, Gasparini F, Manni L (2015) Morphological evidence that the molecularly determined Ciona intestinalis type A and type B are different species: Ciona robusta and Ciona intestinalis. J Zool Syst Evol Res 53(3):186–193. CrossRefGoogle Scholar
  2. Caputi L, Andreakis N, Mastrototaro F, Cirino P, Vassillo M, Sordino P (2007) Cryptic speciation in a model invertebrate chordate. Proc Natl Acad Sci U S A 104(22):9364–9369. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Hikosaka A, Kusakabe T, Satoh N, Makabe KW (1992) Introduction and expression of recombinant genes in ascidian embryos. Develop Growth Differ 34(6):627–634. CrossRefGoogle Scholar
  4. Hikosaka A, Kusakabe T, Satoh N (1994) Short upstream sequences associated with the muscle-specific expression of an actin gene in ascidian embryos. Dev Biol 166(2):763–769. CrossRefPubMedGoogle Scholar
  5. Nydam ML, Harrison RG (2007) Genealogical relationships within and among shallow-water Ciona species (Ascidiacea). Mar Biol 151(5):1839–1847. CrossRefGoogle Scholar
  6. Nydam ML, Harrison RG (2011) Introgression despite substantial divergence in a broadcast spawning marine invertebrate. Evolution 65(2):429–442. CrossRefPubMedGoogle Scholar
  7. Sato A, Shimeld SM, Bishop JDD (2014) Symmetrical reproductive compatibility of two species in the Ciona intestinalis (Ascidiacea) species complex, a model for marine genomics and developmental biology. Zool Sci 31(6):369–274. CrossRefPubMedGoogle Scholar
  8. Satou Y, Satoh N (1996) Two cis-regulatory elements are essential for the muscle-specific expression of an actin gene in the ascidian embryo. Develop Growth Differ 38(5):565–573. CrossRefGoogle Scholar
  9. Satou Y, Imai K, Satoh N (2001) Action of morpholinos in Ciona embryos. Genesis 30(3):103–106. CrossRefPubMedGoogle Scholar
  10. Suzuki MM, Nishikawa T, Bird A (2005) Genomic approaches reveal unexpected genetic divergence within Ciona intestinalis. J Mol Evol 61(5):627–635. CrossRefPubMedGoogle Scholar
  11. Yoshida K, Saiga H (2011) Repression of Rx gene on the left side of the sensory vesicle by Nodal signaling is crucial for right-sided formation of the ocellus photoreceptor in the development of Ciona intestinalis. Dev Biol 354(1):144–150. CrossRefPubMedGoogle Scholar

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© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  1. 1.Department of Zoology, Graduate School of ScienceKyoto UniversitySakyo-ku, KyotoJapan

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