Abstract
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.
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References
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. https://doi.org/10.1111/jzs.12101
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. https://doi.org/10.1073/pnas.0610158104
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. https://doi.org/10.1111/j.1440-169X.1992.tb00031.x
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. https://doi.org/10.1006/dbio.1994.1354
Nydam ML, Harrison RG (2007) Genealogical relationships within and among shallow-water Ciona species (Ascidiacea). Mar Biol 151(5):1839–1847. https://doi.org/10.1007/s00227-007-0617-0
Nydam ML, Harrison RG (2011) Introgression despite substantial divergence in a broadcast spawning marine invertebrate. Evolution 65(2):429–442. https://doi.org/10.1111/j.1558-5646.2010.01153.x
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. https://doi.org/10.2108/zs130249
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. https://doi.org/10.1046/j.1440-169X.1996.t01-1-00013.x
Satou Y, Imai K, Satoh N (2001) Action of morpholinos in Ciona embryos. Genesis 30(3):103–106. https://doi.org/10.1002/gene.1040
Suzuki MM, Nishikawa T, Bird A (2005) Genomic approaches reveal unexpected genetic divergence within Ciona intestinalis. J Mol Evol 61(5):627–635. https://doi.org/10.1007/s00239-005-0009-3
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. https://doi.org/10.1016/j.ydbio.2011.03.006
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Kobayashi, K., Satou, Y. (2018). Microinjection of Exogenous Nucleic Acids into Eggs: Ciona Species. In: Sasakura, Y. (eds) Transgenic Ascidians . Advances in Experimental Medicine and Biology, vol 1029. Springer, Singapore. https://doi.org/10.1007/978-981-10-7545-2_2
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DOI: https://doi.org/10.1007/978-981-10-7545-2_2
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