Difference between revisions of "Team:Vilnius-Lithuania/Basic Part"
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| + | <h3 class="text-heading">Basic part</h3> | ||
| + | <p>In this page we describe our favorite part which is I-F type CRISPR-Cas Cascade complex. It can be found in Parts Registry - <a 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> | ||
| + | <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 (BBa_K1773003) degrades DNA.</p> | ||
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| + | <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 : Csy1, Csy2, Csy3 and Cas6f, with stoichiometry of 1:1:6:1 respectively. Also it has a crRNA molecule, witch 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 Aggregatibacter actinomycetemcomitans 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 (BBa_K1773003) is recruited for DNA degradation.</p> | ||
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Revision as of 10:11, 18 September 2015
Basic part
In this page we describe our favorite part which is I-F type CRISPR-Cas Cascade complex. It can be found in Parts Registry - BBa_K1773005
I-F type CRISPR-Cas Cascade complex
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 (BBa_K1773003) degrades DNA.
Usage and Biology
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, witch 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 Aggregatibacter actinomycetemcomitans 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 (BBa_K1773003) is recruited for DNA degradation.