Team:Freiburg/Highlights
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We developed an ELISA based method. With this method we can quantify the <b>binding efficiency </b>of our proteins. We called this binding assay <b>uniBAss</b>. It is | We developed an ELISA based method. With this method we can quantify the <b>binding efficiency </b>of our proteins. We called this binding assay <b>uniBAss</b>. It is | ||
- | a powerful tool for characterizing the modified dCas9 by assessing | + | a powerful tool for characterizing the modified dCas9 by assessing its DNA binding capacity. |
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Revision as of 15:13, 2 October 2013
HIGHLIGHTS
In the last months we were able to...
- ...constuct a catalytically inactive version of Cas9 and this way generate a new class of DNA binding proteins.
- ...combine this modified dCas9 with different transcriptional effectors.
- ...express this system in various mammalian cell lines.
- ...control human gene expression via our modified CRISPR/Cas system.
- ...regulate gene expression on light stimulus.
- ...make our dCas9 accesible to the whole iGEM comunity by mutating illegal iGEM restriction sites.
In summary we build up a universal toolkit for gene regulation.
6 opportunities to customize your experiments We provide 3 different effectors, 2 methods and 1 effector controler that allows either to effectively repress or activate genes - also on stimulus. Use our custom-tailored Manual Tool to generate individual manuals. Best of all: Its all open source and in iGEM standard! |
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Cas9 makes it possible! First derived from bacterial immune system this wonderful enzyme is a real multifunctional talent. Specific targeting of DNA has never been easier! |
Activation The activation domain VP16 is able to activate transcription of genes. |
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Repression The fusion of the transcriptional repressor domain KRAB leads to synthetic repression of gene expression. |
Chromatin modification (Repression) Specific chromatin modification was achieved by fusing a histone methyltransferase G9a to dCas9. With this protein we are able to specifically repress endogenous gene expression. |
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uniBAss - Binding Assay We developed an ELISA based method. With this method we can quantify the binding efficiency of our proteins. We called this binding assay uniBAss. This is a powerful tool for the characterizing the modified dCas9 by assessing it´s DNA binding capacity. |