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Team:Freiburg bioware
From 2009.igem.org
Project Abstract
After the second place in last year’s iGEM competition we got high expectations and look forward to this year’s Jamboree! We’re working on the development of an universal restriction enzyme to facilitate labwork and enable new techniques.
Restriction enzymes are proteins capable of cutting double or single 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.
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 3×10^9 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 16 bp of length and which is programmable for many different target sequences.
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