Difference between revisions of "Team:Vilnius-Lithuania/Composite Part"

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<h2> Composite Parts</h2>
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        <h3 class="text-heading">Composite part</h3>
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<p>In this page we describe our favorite composite part which is I-F type Cascade complex of I-F type CRISPR-Cas system It can be found in Parts Registry - <a class="clink" href="http://parts.igem.org/Part:BBa_K1773022">BBa_K1773022</a>.</p>
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<h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">Cascade with pLux/cI right promoter and strong RBS</h3>
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<p>I-F type Cascade complex of I-F type CRISPR-Cas system. The Cascade complex distinguishes target DNA and recruits Cas3 (BBa_K1773003) protein, which degrades the target DNA. Cascade complex is comprised of four proteins: Csy1, Csy2, Csy3 and Cas6f. crRNA region coding crRNAs is necessary for complete Cascade complex.
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<h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">Usage and Biology</h3>
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<p>Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems, because it distinguishes foreign DNA and promotes its degradation. This ribonucleoprotein complex is coded by operon of 4 genes: Csy1, Csy2, Csy3 and Cas6f, with stoichiometry of 1:1:6:1 respectively. Also it has a crRNA molecule, which acts like a guide, that brings the whole complex to the target dsDNA sequence. Each Cascade complex has a unique Protospacer Adjacent Motive (PAM) recognition site. The recognizable PAM sequence of this specific I-F Cascade complex from Aggregatibacter actinomycetemcomitans recognizes CC nucleotides directly upstream of the protospacer (the target sequence). In other words, a target sequence needs to have a CC pair on the non-complementary strand of DNA -2 -1 positions of the protospacer. When Cascade complex finds and recognizes the DNA target, then Cas3 (<a class="clink" href="http://parts.igem.org/Part:BBa_K1773003">BBa_K1773003</a>) is recruited for DNA degradation.</p>
  
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<h4>Note</h4>
 
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<p>In order to be considered for the <a href="https://2015.igem.org/Judging/Awards#SpecialPrizes">Best New Composite Part award</a>, you must fill out this page. Please give links to the Registry entries for the Composite parts you have made. Please see the Registry's <a href="http://parts.igem.org/Help:Parts#Basic_and_Composite_Parts"> Help:Parts page</a> for more information on part types.</p>
 
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A composite part is a functional unit of DNA consisting of two or more basic parts assembled together. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_I13507">BBa_I13507</a> is an example of a composite part, consisting of an RBS, a protein coding region for a red fluorescent protein, and a terminator.
 
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<p>New composite BioBrick devices can be made by combining existing BioBrick Parts (like Inverters, Amplifiers, Smell Generators, Protein Balloon Generators, Senders, Receivers, Actuators, and so on).</p>
 
  
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Latest revision as of 10:36, 18 September 2015

Composite part

In this page we describe our favorite composite part which is I-F type Cascade complex of I-F type CRISPR-Cas system It can be found in Parts Registry - BBa_K1773022.

Cascade with pLux/cI right promoter and strong RBS

I-F type Cascade complex of I-F type CRISPR-Cas system. The Cascade complex distinguishes target DNA and recruits Cas3 (BBa_K1773003) protein, which degrades the target DNA. Cascade complex is comprised of four proteins: Csy1, Csy2, Csy3 and Cas6f. crRNA region coding crRNAs is necessary for complete Cascade complex.

Usage and Biology

Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems, because it distinguishes foreign DNA and promotes its degradation. This ribonucleoprotein complex is coded by operon of 4 genes: Csy1, Csy2, Csy3 and Cas6f, with stoichiometry of 1:1:6:1 respectively. Also it has a crRNA molecule, which acts like a guide, that brings the whole complex to the target dsDNA sequence. Each Cascade complex has a unique Protospacer Adjacent Motive (PAM) recognition site. The recognizable PAM sequence of this specific I-F Cascade complex from Aggregatibacter actinomycetemcomitans recognizes CC nucleotides directly upstream of the protospacer (the target sequence). In other words, a target sequence needs to have a CC pair on the non-complementary strand of DNA -2 -1 positions of the protospacer. When Cascade complex finds and recognizes the DNA target, then Cas3 (BBa_K1773003) is recruited for DNA degradation.