Abstract
Functional studies on echinoderms have been reduced to the use of pharmacological treatments. The ability to modulate the genetic expression of regenerating tissues can elucidate potential effectors during this process. Here we describe an effective transfection protocol that allows the introduction of Dicer-substrate interference RNAs (DsiRNAs) for the modulation of gene expression and its characterization during regeneration.
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References
Hannon GJ (2002) RNA interference. Nature 418:244–251
Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Elbashir SM, Lendeckel W, Tuschl T (2001) RNA interference is mediated by 21- and 22- nucleotide RNAs. Genes Dev 15:188–200
Meister G, Tuschi T (2004) Mechanisms of gene silencing by double-stranded RNA. Nature 431:343–349
Kim DH, Behlke MA, Rose SD, Chang MS, Choi S, Rossi JJ (2005) Synthetic dsRNA dicer substrates enhance RNAi potency and efficacy. Nat Biotechnol 23:222–226
Bogarad L, Arnone MI, Chang C, Davidson EH (1998) Interference with gene regulation in living sea urchin embryos: transcription factor knock out (TKO), a genetically controlled vector for blockade of specific transcription factor. PNAS 25:14827–14832
Mashanov VS, Zueva OR, García-Arrarás JE (2015) Myc regulated programmed-cell death and radial glia dedifferentiation after neural injury in an echinoderm. BMC Dev Biol 15:1–9
Bello SA, Abreu-Irizarry RJ, García-Arrarás JE (2015) Primary cell cultures of regenerating holothurian tissues. Methods Mol Biol 1189:283–297
Schacher S, Proshansky E (1983) Neurite regeneration by Aplysia neurons in dissociated cell culture: modulation by Aplysia hemolymph and the presence of the initial axonal segment. J Neurosci 3:2403–2413
Naito Y, Yoshimura J, Morishita S, Ui-Tei K (2009) siDirect 2.0: updated software for designing functional siRNA with reduced seed-dependent off-target effect. BMC Bioinformatics 10:392
Alicea-Delgado M (2019) Functional role of β-catenin and Myc as active players in the canonical Wnt signaling pathway during intestinal regeneration of the sea cucumber Holothuria glaberrima. Thesis, University of Puerto Rico – Río Piedras Campus
Acknowledgments
This project was supported by NIH (Grants R15GM124595 and R21AG057974), and the University of Puerto Rico.
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Alicea-Delgado, M., Bello-Melo, S.A., García-Arrarás, J.E. (2021). RNA Interference on Regenerating Holothurian Gut Tissues. 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_15
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DOI: https://doi.org/10.1007/978-1-0716-0974-3_15
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0973-6
Online ISBN: 978-1-0716-0974-3
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