Our project idea came up in April, when we decided to build a methane biosensor. The aim was clear, as this tool - if we are succesful - can help to solve both an environmental issue and a social problem at once. Nevertheless the method was still unclear: firstly we planned RNA-silencing or using CRISPR. Fortunately at the early stage became clear that transforming methanotrophs is quite complex process. We have to say thank to Lori Giver, researcher at Calysta Inc., who advised us to use the http://methanotroph.org page where we could find many useful information to design our project carefully.
Meanwhile we tried to find help in abroad, it became clear at a certain moment that some researchers working in Department of Biotechnology at the local University of Szeged formerly dealt with a similar issue. Head of Department, Prof. Gabor Rakhely spent a lot of time to make us understand the possible processes of genetic modification of these type of bacteria. Conversations with him lead to the decision to leave both iRNA and CRISPR methods, not only because of the technical difficulties they cause, but also the safety issues arising when somebody use our final product. The stability of a genetic modification in a product which is handled by non-professionals is a key issue for us. Neither the global warming experts who can use our final products at boreal swamps measuring the methane leaking, nor an everyday home-made biogas plant owner in a developing country are trained in using a possibly hazardous material. For this reason we should ensure their personal and also the environmental safety at maximum level. This is why we have chosen finally the triparental mating as a genetic modification method.
Getting this idea we went further in two directions. Firstly we tried to find an excellent expert of the chosen method, triparental mating. All the references we found showed inevitably that one of them is Sergey Stolyar, Associate Professor at University of Idaho. Unfortunately his most important article was unavailable through the resources of the University Library, but we found that there is a Russian language version of his study. We received the photocopy of this article through the courtesy of Prof. Alexander M. Rubtsov (School of Biology at Lomonosov Moscow State University) as he has a collaboration with our PI, Sandor Ban. Finally we decided to contact directly Prof. Stolyar for further details. He was enough kind to respond us and in his email he wrote the appropriate procedure in very details, which helped us to prepare the last step in our project.
At near the final stage when we contacted Anindya Sinha, Professor at National Institute of Advanced Studies at Indian Institute of Science Campus (Bangalore, India), who is an expert in both the contemporary issues in developing countries problems and also in many environmental issues. He advised us to carry on with our project which - as he says - an outstanding example of using up-to-date scientific methods in order to solve urgent issues. He offered the possibility to continue the conversation in the near future. He will help us to gain data on the requirements of a biogas leaking device. The details and prove of the above are as follows:
Lori Giver, Ph.D.
She is the vice-president of Biological Engineering at the Calysta Inc. Calysta is a biotechnological company working with many kinds of bacteria, e.g. Methylococcus capsulatus. We wrote a letter to Professor Giver asking for her help to the project. She was very kind and interested in what we were going to do.
Sergey Stolyar, Ph.D.
He is the Research Associate Professor of Department of Biological Sciences, University of Idaho. He wrote many articles, and was referenced by even more articles, about the transformation of Methylococcus capsulatus. Since we could find the original version of his article only in Russian, we asked him whether he could help us by sending an English translation of the article. He was really helpful, and he not only attached an English translation but he summarized the key steps of making manipulations with methanotrophs as well.
Professor Anindya Sinha, Ph.D.
Professor Sinha's main research area is examining the life of animal societies, such as elephants and primates. His articles and books suggests viewpoints for comparing animal and human societies. He is committed to global envirometal issues and also the concerns of the developing countries in Asia. He is also an expert in managing projects for talented young people interested in Biology.