Team:Freiburg bioware

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Revision as of 08:35, 28 September 2009

FREiGEM 2009

Welcome

Team Freiburg Bioware
The first German team ever to participate in iGEM is back again and after last year's second place we're highly motivated to make some good piece of synthetic biology in 2009. In this year we want to create an universal restriction enzyme to facilitate labwork and enable new techniques.

We're looking forward to meet you on this year's jamboree!

Project Summary

Universal Endonuclease – Cutting Edge Technology

Gene technology is driven by the use of restriction endonucleases. Yet, constraints of limited sequence length and variation recognized by available restriction enzymes pose a major roadblock for synthetic biology. We developed the basis for universal restriction enzymes, primarily for routine cloning but also with potential for in vivo applications. We use a nucleotide cleavage domain fused to a binding domain, which recognizes a programmable adapter that mediates binding to DNA and thus cleavage. As adapter we use readily available modified oligonucleotides, as binding domain anticalins and as cleavage domain FokI moieties engineered for heterodimerization and activity. For cloning, this universal enzyme has merely to be mixed with the sequence specific oligonucleotide and the target DNA. Binding and release are addressed with thermocycling. Additionally, we provide concepts for in vivo applications by external adapter delivery, activity regulation by photo-switching, as well as for modifying an argonaute protein towards a DNA endonuclease.

FREiGEM 2009

The Team
This year our team consists of 18 undergraduate students and 4 advisors.
Read more...

Bioss
We want to thank our main sponsor Bioss for supporting our project.
Read more...

Visitor Locations

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-<p>Restriction enzymes are proteins capable of cutting double or
+<p><b>Universal Endonuclease – Cutting Edge Technology</b></p>
-single
+<p>Gene technology is driven by the use of restriction endonucleases. Yet, constraints of limited sequence length and variation recognized by available restriction enzymes pose a major roadblock for synthetic biology. We developed the basis for universal restriction enzymes, primarily for routine cloning but also with potential for in vivo applications. We use a nucleotide cleavage domain fused to a binding domain, which recognizes a programmable adapter that mediates binding to DNA and thus cleavage. As adapter we use readily available modified oligonucleotides, as binding domain anticalins and as cleavage domain FokI moieties engineered for heterodimerization and activity. For cloning, this universal enzyme has merely to be mixed with the sequence specific oligonucleotide and the target DNA. Binding and release are addressed with thermocycling. Additionally, we provide concepts for in vivo applications by external adapter delivery, activity regulation by photo-switching, as well as for modifying an argonaute protein towards a DNA endonuclease.
-strand nuclein acids. We focus on type II and III restriction enzymes
-which at first bind the DNA strand, then recognize a certain sequence
-pattern where they cut the DNA backbone. There are thousands of
-different restriction enzymes, each with particular recognition and
-cutting sites. These enzymes are essential tools for gene cloning and
-protein expression experiments. Every medical and biological laboratory
-needs to keep dozens of different restriction enzymes in stock for
-regular use. Acquiring and handling so many enzymes is very expensive
-and time consuming. We are determined to simplify this procedure
-totally, by creating one enzyme for every occasion. <br />
-<br />
-One of the biggest challenges of today is to cure diseases by means of
-gene therapy. The aim here is to artifically introduce genetic
-information in somatic cells to substitute DNA-sequences which may
-allow the correction of mutated genes. Particularly in monogenetic
-diseases this would lead to a change of the phenotype. The human genome
-contains 3&times;10^9 bp which code for approximately 30.000
-different genes.
-Alone one single mutation can cause changes which may lead to diseases
-or even death. Because of this it is tried in gene therapy to address
-exactly these mutations. But this means that it is necessary to cause
-specifically on that point a change e.g. by cutting. Restriction
-enzymes could be used as high specific tools for this. But these
-enzymes would have to recognize a sequence of at least 16 bp in oder to
-cut only once in the human genome (416 bp = 4.3*10^9 bp). Most of the
-known 3500 restriction enzymes only recognize sequences with a length
-of 4-8 bp. One application of our work could be to construct an
-artificial restriction enzyme with a recognition sequence of at least
-bp of length and which is programmable for many different target
-sequences.
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