Team:TCU Taiwan/Parts

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BBa_K1473005

Phagemid pBluescript is a special plasmid, it functions as a normal plasmid when transformed into bacteria. But when helper phage M13KO7 infect the bacteria, it will produce progeny M13 phage as vector for pBluescript.

pBluescript contains a f1 ori while M13KO7 helper phage’s f1 ori has been knock-down. So after the infection of M13KO7, its genome can produce protein and replicate but these proteins will package pBluescript as their true “genome”. As a result, the progeny phage’s genome will only contains pBluescript’s MCS, they cannot make next generation.

Fig.1

BBa_K1473009

Here we combined a CRISPR system(BBa_K1218011) with an inducible promoter(BBa_K914003). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch if on or off.

Source :Science

Parts Sandbox

Click on the name of the parts for detailed information.

Name	Type	Description	Designer	Length
BBa_K1473001	DNA	gRNA 1	Shan, Zhe	22
BBa_K1473002	DNA	gRNA 2	Shan, Zhe	22
BBa_K1473003	DNA	gRNA 3	Shan, Zhe	22
BBa_K1473004	DNA	gRNA 4	Shan, Zhe	22
BBa_K1473005	Plasmid_Backbone	phagemid pBluescript	Shan, Zhe	2929
BBa_K1473006	DNA	gRNA for ampR 1	Shan, Zhe	22
BBa_K1473007	DNA	gRNA for ampR 2	Shan, Zhe	22
BBa_K1473008	DNA	gRNA for ampR 3	Shan, Zhe	22
BBa_K1473009	Coding	pRha induced CRISPR	Shan, Zhe	5011
BBa_K1473010	DNA	gRNA for ampR 4	Shan, Zhe	22
BBa_K1473011	DNA	gRNA for ampR 5	Shan, Zhe	22
BBa_K1473012	DNA	gRNA for NeoR/kanR 1	Shan, Zhe	22
BBa_K1473013	DNA	gRNA for NeoR/kanR 2	Shan, Zhe	22
BBa_K1473014	DNA	gRNA for NeoR/kanR 3	Shan, Zhe	22
BBa_K1473015	DNA	gRNA for NeoR/kanR 4	Shan, Zhe	22
BBa_K1473016	DNA	gRNA for NeoR/kanR 5	Shan, Zhe	22
BBa_K1473017	DNA	gRNA for tetR 1	Shan, Zhe	22
BBa_K1473018	DNA	gRNA for tetR 2	Shan, Zhe	22
BBa_K1473019	DNA	gRNA for tetR 3	Shan, Zhe	22
BBa_K1473020	DNA	gRNA for tetR 4	Shan, Zhe	22
BBa_K1473021	DNA	gRNA for tetR 5	Shan, Zhe	22

TCU_Taiwan 2014 iGEM Team Parts

gRNA

Fig.2

Fig.3

We put a sacI restriction cutting site downstream to gRNA (not shown in sequence information) and ligated it back to pSB1C3 with correct prefix and surffix. Then we cut this recombinant plasmid with sacI and run gel electrophoresis for testing. As we can see in this image, because pSB1C3 only contains 1 sacI cutting site itself, so it would not be cut into 2 parts when no gRNA is ligated inside. But if there is, then sacI will cut this recombinant plasmid into 2 parts, whose lengths are 939 bp and 1159 bp. Obviously, we have successfully synthesized this gRNA.

Parts Table

This table is from the Database of iGEM.

Any parts our team has created will appear in this table below:

<groupparts>iGEM014 TCU_Taiwan</groupparts>

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Team Members	Project	Parts	Human Pratics	Modeling	Safety	Notebook	Attributions

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@@ Line 235: / Line 235: @@
 </tr>
 <tr>
-   <td valign="top"><font size="3" face="Verdana" color="#333">Here we combined a CRISPR system(BBa_K1218011) with an inducible promoter(BBa_K914003). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal
+   <td valign="top"><font size="3" face="Verdana" color="#333">Here we combined a CRISPR system(<a href="http://parts.igem.org/Part:BBa_K1218011" target="_blank">BBa_K1218011</a>) with an inducible promoter(<a href="http://parts.igem.org/Part:BBa_K1218011" target="_blank">BBa_K914003</a>). The CRISPR system contains a tracrRNA , a Cas9 protein and a minimal
      CRISPR array, it functions to knockout target gene. We give this system an inducible promoter so we can decided when to switch if on or off. </font></td>
      <td width="45%" align="center"><table width="100%" border="0" cellspacing="0" cellpadding="0">