2019 Echinoderm Protocols - Methods in Cell Biology and CRISPR/Cas in Echinoderms: Difference between pages

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== Methods in Cell Biology 2019==
Welcome to the Echinobase CRISPR/Cas resource. A brief literature and method review is followed by tables of gRNA spacer sequences.


<p>In 2019, two volumes of Echinoderm protocols were published in Methods in Cell Biology.</p>
Updated December 2020
<p>These volumes were edited by [https://www.echinobase.org/community/person.do?method=display&personId=4063&tabId=0 Amro Hamdoun] and [https://www.echinobase.org/community/person.do?method=display&personId=4062 Kathy Foltz].</p>




<p>[https://www.sciencedirect.com/bookseries/methods-in-cell-biology/vol/150/suppl/C '''Volume 150''']</p>
'''''S. purpuratus'' genome editing to create insertions and deletions'''


<p>Section I: Procurement and culturing of established and emerging echinoderm models</p>
To date CRISPR/Cas9 has been used to introduce insertion and deletion mutations (indels) into ''S. purpuratus nodall'' ([https://www.echinobase.org/literature/article.do?method=display&articleId=44372 Lin and Su 2016]), ''polyketide synthase 1'', ''gcml'' (Oulhen and Wessel 2016), ''nanos2l'' ([https://www.echinobase.org/literature/article.do?method=display&articleId=45207 Oulhen et al. 2017]) and ''dll1'' (''delta'') ([https://www.echinobase.org/literature/article.do?method=display&articleId=45720 Mellott et al. 2017]) genes. Attempts to mutate ''foxy'' (Oulhen et al. 2019) were unsuccessful. A number of different methods were used for gRNA synthesis (several using pT7-gRNA)  and NLS-SpCas9-NLS (pCS2-nCas9n (zebrafish codon-optimized), or pCS2-3xFLAG-NLS-SpCas9-NLS (codon optimized for human with a 3XFLAG-tag) were used in these studies (see below for details). The gRNAs and mRNAs were microinjected into fertilized eggs.
<p>Section II: Experimental embryology approaches</p>
<p>Section III: Approaches for assessing environmental influences on adults and embryos</p>
<p>Section IV: Sea urchins in the classroom</p>
 
 
<p>[https://www.sciencedirect.com/bookseries/methods-in-cell-biology/vol/151/suppl/C '''Volume 151''']</p>
 
<p>1: Fertilization</p>
<p>2: Cytokinesis and embryology</p>
<p>3: Genomics and morphogenesis</p>
<p>Section I: Methods for genome and transcriptome analysis</p>
<p>Section II: Genome-editing and proteomics</p>
<p>Section III: Imaging of echinoderm embryos</p>
<p>Section IV: Methods for measurement of intracellular signals in eggs, sperm and embryos</p>

Revision as of 10:15, 1 December 2020

Welcome to the Echinobase CRISPR/Cas resource. A brief literature and method review is followed by tables of gRNA spacer sequences.

Updated December 2020


S. purpuratus genome editing to create insertions and deletions

To date CRISPR/Cas9 has been used to introduce insertion and deletion mutations (indels) into S. purpuratus nodall (Lin and Su 2016), polyketide synthase 1, gcml (Oulhen and Wessel 2016), nanos2l (Oulhen et al. 2017) and dll1 (delta) (Mellott et al. 2017) genes. Attempts to mutate foxy (Oulhen et al. 2019) were unsuccessful. A number of different methods were used for gRNA synthesis (several using pT7-gRNA) and NLS-SpCas9-NLS (pCS2-nCas9n (zebrafish codon-optimized), or pCS2-3xFLAG-NLS-SpCas9-NLS (codon optimized for human with a 3XFLAG-tag) were used in these studies (see below for details). The gRNAs and mRNAs were microinjected into fertilized eggs.