Difference between revisions of "Team:Vilnius-Lithuania/Basic Part"
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<p>In this page we describe our favorite basic part, which is I-F type CRISPR-Cas Cascade complex. It can be found in Parts Registry - <a class="clink" href="http://parts.igem.org/Part:BBa_K1773005">BBa_K1773005</a>.</p> | <p>In this page we describe our favorite basic part, which is I-F type CRISPR-Cas Cascade complex. It can be found in Parts Registry - <a class="clink" href="http://parts.igem.org/Part:BBa_K1773005">BBa_K1773005</a>.</p> | ||
<h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">I-F type CRISPR-Cas Cascade complex</h3> | <h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">I-F type CRISPR-Cas Cascade complex</h3> | ||
− | <p>I-F type CRISPR-Cas Cascade complex, which has a mutated EcoRI restriction site to make it biobrick compatible. This protein complex needs to be coexpressed with a crRNA region | + | <p>I-F type CRISPR-Cas Cascade complex, which has a mutated EcoRI restriction site to make it biobrick compatible. This protein complex needs to be coexpressed with a crRNA region to make a functional Cascade ribonucleoprotein complex. Together with Cas3 (<a class="clink" href="">BBa_K1773003</a>) degrades DNA.</p> |
<h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">Usage and Biology</h3> | <h3 style="border-left: 5px solid rgb(236,151,31); padding-left: 5px; border-bottom: none">Usage and Biology</h3> | ||
− | <p>Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems. This ribonucleoprotein complex is coded by operon of 4 genes | + | <p>Cascade (CRISPR Associated Complex for Antiviral Defence) complex is an essential component of I-type CRISPR-Cas systems. 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 recognisable PAM sequence of this specific I-F Cascade complex from <em>Aggregatibacter actinomycetemcomitans</em> recognises 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 recognises the DNA target, then Cas3 (<a class="clink" href= "">BBa_K1773003</a>) is recruited for DNA degradation.</p> |
Revision as of 10:32, 18 September 2015