Talk:Sea Urchin Development
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Cite this page: Hill, M.A. (2020, January 26) Embryology Sea Urchin Development. Retrieved from https://embryology.med.unsw.edu.au/embryology/index.php/Talk:Sea_Urchin_Development
10 Most Recent Papers
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Sea Urchin Development
<pubmed limit=5>Sea Urchin Development</pubmed>
Sea Urchin Embryology
<pubmed limit=5>Sea Urchin Embryology</pubmed>
An optogenetic approach to control protein localization during embryogenesis of the sea urchin
Dev Biol. 2018 Sep 1;441(1):19-30. doi: 10.1016/j.ydbio.2018.06.015. Epub 2018 Jun 26.
Uchida A1, Yajima M2. Author information Abstract Light inducible protein-protein interactions have been used to manipulate protein localization and function in the cell with utmost spatial and temporal precision. In this technical report, we use a recently developed optogenetic approach to manipulate protein localization in the developing sea urchin embryo. A photosensitive LOV domain from Avena sativa phototropin1 cages a small peptide that binds the engineered PDZ domain (ePDZ) upon blue light irradiation. Using this system, mCherry tagged proteins fused with the LOV domain were recruited to ectopic sub-cellular regions such as the membrane, microtubules, or actin by GFP tagged proteins fused with the ePDZ domain upon blue light irradiation within 1-3 min in the sea urchin embryo. The efficiency and speed of recruitment of each protein to its respective subcellular region appeared to be dependent on the power and duration of laser irradiation, as well as the respective level of affinity to the tagged location. Controlled laser irradiation allowed partial recruitment of the spindle to the membrane, and resulted in cell blebbing. Vasa, a cell cycle and germline factor that localizes on the spindle and enriches in the micromeres at 8-16 cell stage was recruited to ectopic sites, preventing normal enrichment. Continuous blue light activation with a regular blue aquarium light over two days of culture successfully induced LOV-ePDZ binding in the developing embryos, resulting in continued ectopic recruitment of Vasa and failure in gastrulation at Day 2. Although some cytotoxicity was observed with prolonged blue light irradiation, this optogenetic system provides a promising approach to test the sub-cellular activities of developmental factors, as well as to alter protein localization and development during embryogenesis. KEYWORDS: Embryo; Live imaging; Optogenetics; Sea urchin PMID: 29958898 PMCID: PMC6089079 [Available on 2019-09-01] DOI: 10.1016/j.ydbio.2018.06.015
Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo
Dev Dyn. 2017 Dec;246(12):1036-1046. doi: 10.1002/dvdy.24586. Epub 2017 Oct 13.
Shevidi S1, Uchida A1, Schudrowitz N1, Wessel GM1, Yajima M1. Author information Abstract BACKGROUND: A single base pair mutation in the genome can result in many congenital disorders in humans. The recent gene editing approach using CRISPR/Cas9 has rapidly become a powerful tool to replicate or repair such mutations in the genome. These approaches rely on cleaving DNA, while presenting unexpected risks. RESULTS: In this study, we demonstrate a modified CRISPR/Cas9 system fused to cytosine deaminase (Cas9-DA), which induces a single nucleotide conversion in the genome. Cas9-DA was introduced into sea urchin eggs with sgRNAs targeted for SpAlx1, SpDsh, or SpPks, each of which is critical for skeletogenesis, embryonic axis formation, or pigment formation, respectively. We found that both Cas9 and Cas9-DA edit the genome, and cause predicted phenotypic changes at a similar efficiency. Cas9, however, resulted in significant deletions in the genome centered on the gRNA target sequence, whereas Cas9-DA resulted in single or double nucleotide editing of C to T conversions within the gRNA target sequence. CONCLUSIONS: These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations. Developmental Dynamics 246:1036-1046, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc. KEYWORDS: CRISPR; Cas9; deaminase; genome editing; sea urchin PMID: 28857338 PMCID: PMC5872153 DOI: 10.1002/dvdy.24586