Microinjection of Exogenous Nucleic Acids into Eggs: Ciona Species
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.
KeywordsDevelopmental biology Microinjection Ascidian Ciona Nucleic acid DNA RNA Morpholino oligonucleotide Egg Embryo
- 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 CrossRefGoogle Scholar
- 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 CrossRefGoogle Scholar
- 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 CrossRefGoogle Scholar
- 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 CrossRefPubMedGoogle Scholar