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| | __NOTOC__ | | __NOTOC__ |
| | <partinfo>BBa_K2926003 short</partinfo> | | <partinfo>BBa_K2926003 short</partinfo> |
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| | + | Long description |
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| − | Cas 13a Lsh
| + | <!-- Add more about the biology of this part here |
| − | This
| + | ===Usage and Biology=== |
| − | <i>Leptotcichia shahii</i> | + | |
| − | | + | |
| − | For the full characterization of this part, see BBa (Lbu)
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| − | | + | |
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| − | Cas13a Lwa
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| − | <i>Leptotrichia wadei</i>
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| − | For the full characterization of this part, see BBa (Lbu)
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| − | <h1>Cas 13a Lbu</h1>
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| − | ==Usage and Biology== | + | |
| − | <html>
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| − | <p>
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| − | Cas13a from <i>Leptorichia bucallis</i> (Lbu) is a single-effector RNA guided ribonuclease.
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| − | Upon recognition of its RNA target the activated Cas13a performs RNA cleavage of nearby non-target RNAs.
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| − | This is called "colleteral" RNA cleavage and can lead to cell death in prokaryotes.(Liu et al., 2017)
| + | |
| − | CRISPR (clustered regulary interspaced short palindormic repeat) is an
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| − | adaptive immune systems that provides protection against vaious threats to the cell.
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| − | Cas13a is part of a CRISPR-Cas system which protects the cell against mobile genetic elemens like viruses,
| + | |
| − | transposable elements and conjugative palsmids. (Koonin and Zhang, 2017)
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| − | So far different verions of the protein where found in <i>Leptorichia bucallis</i>, <i>Leptotrichia wadei</i> and <i>Leptotcichia shahii</i>. (Gootenberg et al., 2017)
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| − | The CRISPR cluster, containing the guide RNAs for Cas13a is transciped as pre-CRISPR RNA (pre-crRNA) and processed into crRNA.
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| − | Unlike other Cas effectors, Cas13a is capable of processing the pre-crRNA by itself.
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| − | It is cleaving the pre-crRNA 4 bais pairs upstram of the stem-loop,
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| − | binding the cr-RNA sequence and then building the cas13a:crRNA surveillance complex. (East-seletsky et al., 2017)
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| − | </p>
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| − | [[File:T--Bielefeld-CeBiTec--Background_Cas13a_Background.png|600px|thumb|center|
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| − | <b>As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA.
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| − | Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.
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| − | </b>
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| − | ]]
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| − | | + | |
| − | Grafik zu pre-crRNA processing
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| | + | <!-- --> |
| | <span class='h3bb'>Sequence and Features</span> | | <span class='h3bb'>Sequence and Features</span> |
| | <partinfo>BBa_K2926003 SequenceAndFeatures</partinfo> | | <partinfo>BBa_K2926003 SequenceAndFeatures</partinfo> |
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| − | __TOC__
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| | <!-- Uncomment this to enable Functional Parameter display | | <!-- Uncomment this to enable Functional Parameter display |
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| | <partinfo>BBa_K2926003 parameters</partinfo> | | <partinfo>BBa_K2926003 parameters</partinfo> |
| | <!-- --> | | <!-- --> |
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| − | ==Computational Analysis of Cas13a Lbu==
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| − | <!DOCTYPE html>
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| − | ==Plasmid Design==
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| − | ===Sequencing Results===
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| − | ==Transciption Analyses via (q)RT-PCR==
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| − | ==Protein Purification==
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| − | ===Intein dependtend purification===
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| − | ===SDS PAGE===
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| − | ===MALDI Analyses===
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| − | ===blabla in vitro Assay===
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| − | ==gRNA Modeling???==
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| − | ==Cultivations==
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| − | ===Shake Flask Cultivation===
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| − | ===Micocultivations===
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| − | ==Outlook==
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| − | ==References==
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