CRISPR/Cas in Echinoderms and Sea Urchin Fertilization Laboratory: Difference between pages

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<span class="newwin">[http://new.echinobase.org/literature/article.do?method=display&articleId=42133 '''Sea Urchin Fertilization Laboratory''']</span><br />
Vic Vacquier developed a handout for university students on how to observe sea urchin fertilization in a university lab class setting. All that is needed are sea urchin gametes, a microscope, a few solutions and this handout. The handout works well for both 1 or 2 three hour labs. The student reads through the handout, makes the simple observations, and does the simple, direct experiments to gain insight into the fascinating process of sperm-egg interaction and egg activation.


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


Updated December 2020
<span class="newwin">'''Ocean Life Word Search'''</span><br />


Word searches are fun for students of all ages.
[https://puzzlemaker.discoveryeducation.com/word-search Here is a site] that can be used to make puzzles and a list of terms to get you started.<br />
<p style="margin-left: 20px">MARINE</p>
<p style="margin-left: 20px">ENVIRONMENT</p>
<p style="margin-left: 20px">INVERTEBRATES</p>
<p style="margin-left: 20px">ECHINODERMS</p>
<p style="margin-left: 20px">SEAURCHIN</p>
<p style="margin-left: 20px">SEASTAR</p>
<p style="margin-left: 20px">ANEMONE</p>
<p style="margin-left: 20px">CORAL</p>
<p style="margin-left: 20px">SPONGES</p>
<p style="margin-left: 20px">FISH</p>
<p style="margin-left: 20px">ECHINOBASE</p>
<p style="margin-left: 20px">DEVELOPMENT</p>
<p style="margin-left: 20px">LARVAE</p>
<p style="margin-left: 20px">MEASUREMENT</p><br />


'''''S. purpuratus'' genome editing to create insertions and deletions'''
<span class="newwin">'''Echinoderm Worksheet'''</span><br />


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.
This worksheet can be used as an introduction to echinoderms.<br />
 
<p style="margin-left: 20px">[[File:Echinoderm_Worksheet.jpg|frameless|200px]]</p>
 
<span class="newwin">'''Cut and Paste Echinoderm Life Cycles'''</span><br />
 
This images show the life cycle of the bat star.<br />
 
<p style="margin-left: 20px">[[File:Sea_Star_Life_Cycle.jpg|frameless|200px]]</p>
 
The students then cut and paste sea urchin life cycle stages.<br />

Revision as of 16:03, 19 February 2021

Sea Urchin Fertilization Laboratory
Vic Vacquier developed a handout for university students on how to observe sea urchin fertilization in a university lab class setting. All that is needed are sea urchin gametes, a microscope, a few solutions and this handout. The handout works well for both 1 or 2 three hour labs. The student reads through the handout, makes the simple observations, and does the simple, direct experiments to gain insight into the fascinating process of sperm-egg interaction and egg activation.


Ocean Life Word Search

Word searches are fun for students of all ages. Here is a site that can be used to make puzzles and a list of terms to get you started.

MARINE

ENVIRONMENT

INVERTEBRATES

ECHINODERMS

SEAURCHIN

SEASTAR

ANEMONE

CORAL

SPONGES

FISH

ECHINOBASE

DEVELOPMENT

LARVAE

MEASUREMENT


Echinoderm Worksheet

This worksheet can be used as an introduction to echinoderms.

Cut and Paste Echinoderm Life Cycles

This images show the life cycle of the bat star.

The students then cut and paste sea urchin life cycle stages.