Sea Urchin Development - Revision history

Z8600021 at 12:27, 6 Ocak 2019

2019-01-06T12:27:27Z

← Older revision		Revision as of 23:27, 6 Ocak 2019
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	Search term: [http://www.ncbi.nlm.nih.gov/pubmed/?term=Sea+Urchin+Embryology ''Sea Urchin Embryology'']		Search term: [http://www.ncbi.nlm.nih.gov/pubmed/?term=Sea+Urchin+Embryology ''Sea Urchin Embryology'']

	~~<pubmed limit=5>Sea Urchin Embryology</pubmed>~~
	\|}		\|}
	{\| class="wikitable mw-collapsible mw-collapsed"		{\| class="wikitable mw-collapsible mw-collapsed"
	! Older papers		! Older papers
	\|-		\|-
	\|		\| {{Older papers}}
	* '''Dynamics of Delta/{{Notch}} signaling on endomesoderm segregation in the sea urchin embryo.'''{{#pmid:20023163\|PMID20023163}} "Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/{{Notch}} pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages."		* '''Dynamics of Delta/{{Notch}} signaling on endomesoderm segregation in the sea urchin embryo.'''{{#pmid:20023163\|PMID20023163}} "Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/{{Notch}} pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages."

Z8600021 at 00:58, 25 October 2018

2018-10-25T00:58:01Z

← Older revision		Revision as of 11:58, 25 October 2018
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	* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."		* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."

	* '''Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo'''{{#pmid:28857338\|PMID28857338}} "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.		* '''Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo'''{{#pmid:28857338\|PMID28857338}} "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. These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations."
	These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations."

	* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."		* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."

Z8600021 at 00:28, 25 October 2018

2018-10-25T00:28:40Z

← Older revision		Revision as of 11:28, 25 October 2018
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	\| valign=top\|Spermatozoa Chemotaxis		\| valign=top\|Spermatozoa Chemotaxis
	<br>		<br>
	Sea urchin spermatozoa chemotaxis towards resact.{{#pmid:23183693\|PMID23183693}}		Sea urchin {{spermatozoa}} chemotaxis towards resact.{{#pmid:23183693\|PMID23183693}} Resact causes stimulation of spermatozoa respiration and motility through intracellular alkalinization, transient elevations of cAMP, cGMP and calcium levels in sperm cells, and transient activation and subsequent inactivation of the membrane form of guanylate cyclase.
	<br><br><br>		<br><br><br>
	{{Spermatozoa chemotaxis movie‎}}		{{Spermatozoa chemotaxis movie‎}}

Z8600021 at 00:27, 25 October 2018

2018-10-25T00:27:46Z

← Older revision		Revision as of 11:27, 25 October 2018
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	{\|		{\|
	\| width=~~520px~~\|<html5media height="~~550~~" width="~~512~~">File:Spermatozoa chemotaxis PMID23183693.mp4</html5media>		\| width=310px\|<html5media height="300" width="280">File:Spermatozoa chemotaxis PMID23183693.mp4</html5media>
	\| Sea urchin spermatozoa chemotaxis towards resact.{{#pmid:23183693\|PMID23183693}}		\| valign=top\|Spermatozoa Chemotaxis
			<br>
			Sea urchin spermatozoa chemotaxis towards resact.{{#pmid:23183693\|PMID23183693}}
			<br><br><br>
	{{Spermatozoa chemotaxis movie‎}}		{{Spermatozoa chemotaxis movie‎}}
	\|}		\|}

Z8600021 at 00:26, 25 October 2018

2018-10-25T00:26:20Z

← Older revision		Revision as of 11:26, 25 October 2018
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	\|}		\|}
	==Movies==		==Movies==

	{\|		{\|
	\| Sea urchin spermatozoa chemotaxis towards resact.		\| width=520px\|<html5media height="550" width="512">File:Spermatozoa chemotaxis PMID23183693.mp4</html5media>
	\| {{Spermatozoa chemotaxis movie‎}}		\| Sea urchin spermatozoa chemotaxis towards resact.{{#pmid:23183693\|PMID23183693}}

			{{Spermatozoa chemotaxis movie‎}}
	\|}		\|}

Z8600021 at 00:24, 25 October 2018

2018-10-25T00:24:28Z

← Older revision		Revision as of 11:24, 25 October 2018
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	[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]		[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]

	The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]; [[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig14\|S. Horstadius, 1928]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.		The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]; Lillie, 1912{{#pmid:17735765\|PMID17735765}} [[Spermatozoa Chemotaxis]]; [[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig14\|S. Horstadius, 1928]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.

Z8600021 at 00:20, 25 October 2018

2018-10-25T00:20:51Z

← Older revision		Revision as of 11:20, 25 October 2018
Line 3:		Line 3:
	[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]		[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]

	The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.		The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]; [[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig14\|S. Horstadius, 1928]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.



	[[File:Sea urchin left-right asymmetry.jpg\|600px]]		[[File:Sea urchin left-right asymmetry.jpg\|600px]]


	:'''Links:''' [[:Category:Sea Urchin\|Category:Sea Urchin]]		:'''Links:''' [[:Category:Sea Urchin\|Category:Sea Urchin]]


	~~{{Animals}}~~

	==Some Recent Findings==		==Some Recent Findings==

Z8600021 at 00:18, 25 October 2018

2018-10-25T00:18:22Z

← Older revision		Revision as of 11:18, 25 October 2018
Line 3:		Line 3:
	[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]		[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]

	The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.		The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.

Z8600021 at 00:16, 25 October 2018

2018-10-25T00:16:55Z

← Older revision		Revision as of 11:16, 25 October 2018
Line 3:		Line 3:
	[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]		[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]

	The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. ~~This~~ system has been used recently to study early molecular controls of patterning and axis formation.		The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.

Z8600021 at 00:13, 25 October 2018

2018-10-25T00:13:10Z

← Older revision		Revision as of 11:13, 25 October 2018
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	\|		\|
	* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."		* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."

			* '''Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo'''{{#pmid:28857338\|PMID28857338}} "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.
			These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations."

	* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."		* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."

	* '''Dynamics of Delta/{{Notch}} signaling on endomesoderm segregation in the sea urchin embryo.'''{{#pmid:20023163\|PMID20023163}} "Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/{{Notch}} pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages."
	\|}		\|}
	{\| class="wikitable mw-collapsible mw-collapsed"		{\| class="wikitable mw-collapsible mw-collapsed"
Line 36:		Line 37:
	\|-		\|-
	\|		\|
			* '''Dynamics of Delta/{{Notch}} signaling on endomesoderm segregation in the sea urchin embryo.'''{{#pmid:20023163\|PMID20023163}} "Endomesoderm is the common progenitor of endoderm and mesoderm early in the development of many animals. In the sea urchin embryo, the Delta/{{Notch}} pathway is necessary for the diversification of this tissue, as are two early transcription factors, Gcm and FoxA, which are expressed in mesoderm and endoderm, respectively. Here, we provide a detailed lineage analysis of the cleavages leading to endomesoderm segregation, and examine the expression patterns and the regulatory relationships of three known regulators of this cell fate dichotomy in the context of the lineages."

	* '''The expression and distribution of {{Wnt}} and Wnt receptor mRNAs during early sea urchin development.'''{{#pmid:19853669\|PMID19853669}} "In this study, we identified all Wnt and Wnt receptor mRNAs that are present in unfertilized sea urchin eggs and early embryos and analyzed their distributions along the primary (AV) axis. Our findings indicate that the asymmetric distribution of a maternal Wnt or Wnt receptor mRNA is unlikely to be a primary determinant of the polarized stabilization of beta-catenin along the AV axis. This contrasts sharply with findings in other organisms and points to remarkable evolutionary flexibility in the molecular mechanisms that underlie this otherwise very highly conserved patterning process."		* '''The expression and distribution of {{Wnt}} and Wnt receptor mRNAs during early sea urchin development.'''{{#pmid:19853669\|PMID19853669}} "In this study, we identified all Wnt and Wnt receptor mRNAs that are present in unfertilized sea urchin eggs and early embryos and analyzed their distributions along the primary (AV) axis. Our findings indicate that the asymmetric distribution of a maternal Wnt or Wnt receptor mRNA is unlikely to be a primary determinant of the polarized stabilization of beta-catenin along the AV axis. This contrasts sharply with findings in other organisms and points to remarkable evolutionary flexibility in the molecular mechanisms that underlie this otherwise very highly conserved patterning process."
	\|}		\|}

← Older revision		Revision as of 11:58, 25 October 2018
Line 18:		Line 18:
	* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."		* '''An optogenetic approach to control protein localization during embryogenesis of the sea urchin'''{{#pmid:29958898\|PMID29958898}} "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....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."

	* '''Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo'''{{#pmid:28857338\|PMID28857338}} "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.		* '''Single nucleotide editing without DNA cleavage using CRISPR/Cas9-deaminase in the sea urchin embryo'''{{#pmid:28857338\|PMID28857338}} "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. These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations."
	These results suggest that the Cas9-DA approach may be useful for manipulating gene activity with decreased risks of genomic aberrations."

	* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."		* '''Implication of HpEts in gene regulatory networks responsible for specification of sea urchin skeletogenic primary mesenchyme cells'''{{#pmid:20695779\|PMID20695779}} "The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos."

← Older revision		Revision as of 11:20, 25 October 2018
Line 3:		Line 3:
	[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]		[[File:Sea_urchin_SEM01.jpg\|thumb\|Sea Urchin- 2 cell stage]]

	The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.		The {{sea urchin}} embryo initially undergoes ten cycles of cell division forming a single epithelial layer enveloping a blastocoel, followed by gastrulation producing the three germ layers. Historically, one of the earliest systems used in developmental biology ([[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig05\|H. Driesch, 1892]]; [[Book_-_The_Emergence_of_Experimental_Embryology_in_the_United_States#Studies_on_Epigenesis_the_Physicochemical_Nature_of_Cytoplasm\|J. Loeb, 1893]]; [[Book - The Emergence of Experimental Embryology in the United States#Hilfer1990-Fig14\|S. Horstadius, 1928]]), this embryo system has been used recently to study early molecular controls of patterning and axis formation.



	[[File:Sea urchin left-right asymmetry.jpg\|600px]]		[[File:Sea urchin left-right asymmetry.jpg\|600px]]


	:'''Links:''' [[:Category:Sea Urchin\|Category:Sea Urchin]]		:'''Links:''' [[:Category:Sea Urchin\|Category:Sea Urchin]]


	~~{{Animals}}~~

	==Some Recent Findings==		==Some Recent Findings==