1. Using CRISPR/Cas9 to identify genes required for mechanosensory neuron development and function.
- Author
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Johnson CJ, Kulkarni A, Buxton WJ, Hui TY, Kayastha A, Khoja AA, Leandre J, Mehta VV, Ostrowski L, Pareizs EG, Scotto RL, Vargas V, Vellingiri RM, Verzino G, Vohra R, Wakade SC, Winkeljohn VM, Winkeljohn VM, Rotterman TM, and Stolfi A
- Subjects
- Animals, Larva genetics, Metamorphosis, Biological genetics, Sensory Receptor Cells, CRISPR-Cas Systems, Ciona
- Abstract
Tunicates are marine, non-vertebrate chordates that comprise the sister group to the vertebrates. Most tunicates have a biphasic lifecycle that alternates between a swimming larva and a sessile adult. Recent advances have shed light on the neural basis for the tunicate larva's ability to sense a proper substrate for settlement and initiate metamorphosis. Work in the highly tractable laboratory model tunicate Ciona robusta suggests that sensory neurons embedded in the anterior papillae transduce mechanosensory stimuli to trigger larval tail retraction and initiate the process of metamorphosis. Here, we take advantage of the low-cost and simplicity of Ciona by using tissue-specific CRISPR/Cas9-mediated mutagenesis to screen for genes potentially involved in mechanosensation and metamorphosis, in the context of an undergraduate 'capstone' research course. This small screen revealed at least one gene, Vamp1/2/3, which appears crucial for the ability of the papillae to trigger metamorphosis. We also provide step-by-step protocols and tutorials associated with this course, in the hope that it might be replicated in similar CRISPR-based laboratory courses wherever Ciona are available., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)
- Published
- 2023
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