The Nobel Prize for CRISPR is one of the most exciting ever assigned in chemistry and one of the most celebrated in the media, for reasons related to the invention and the inventors alike. On the one hand, the technique is changing the practice and the image of genetic engineering. On the other hand, Jennifer Doudna and Emmanuelle Charpentier are not merely great scientists; they are a success story in cracking the glass ceiling and a symbol of the strength of collaboration.
Among the genome editing pioneers, Doudna is the one who has put the most effort into initiating a debate on responsible innovation within and outside the labs by gathering working groups and writing for lay readers. Following her example, a community of researchers committed to dialogue has grown around CRISPR.
Recently an online symposium called World CRISPR Day offered the chance to listen to Doudna in the Nobel aftermath. Below you can read some extracts of her opening lecture and Q&A session, and measure the genome editing community’s mood.
“CRISPR allows anyone with a little skills in molecular biology to manipulate genome, and that comes with extraordinary opportunity as well as incredible responsibility; I’ve been involved in both these aspects of the field in the last few years.”
“I want to point out first of all that this technology comes from curiosity-driven research that was conducted by a number of scientists but including myself and Emmanuelle Charpentier. We have been interested in the last 12 years in understanding how CRISPR works as an immune system in bacteria to provide protection against viruses. And it was through studying the molecular mechanisms of this that we understood how a protein called Cas9 works as an RNA-guided DNA cutter, an enzyme that can cut DNA at precise positions defined by the sequence of the guide RNA. With that capability, it was possible to harness it as a technology for genome editing in cells that are able to repair DNA breaks using non-homologous end joining or homology directed repair.”
“The work that Martin Jinek e Krzysztof Chylinski made in our laboratories, they found that Cas9 was a dual RNA guided enzyme, with two separated RNA molecules that provide the targeted information. They showed that these two RNAs could be combined into a single guide format that allowed a simplified system for programming Cas9 in cells, and this was for us that kind of proverbial moment when we realized that a curiosity-driven project to understand bacterial immunity had moved into something really different from what we imagined at the start of the project, which was an idea for a technology that could be used to trigger double-stranded breaks in eukaryotes and any cell type and trigger DNA repair introducing specific genome edits in these cells.”
“Where is all this going? One of the really exciting applications, it is not only one, there are many, is the opportunity to cure genetic diseases. A few years ago it would have sounded like science fiction, but today we are a the point that we can manipulate DNA in cells in the laboratory to correct disease-causing mutations like the one that causes sickle cell disease. Clinical trials are underway using CRISPR to treat this disease, and there is the announcement about a patient that has this disease effectively cured.”
“This is exciting for the field but also brings with it extraordinary challenges in terms of access and making sure that this technology is available to all those around the world who need it. The next stage in this field is how to make sure that people who need access to this technology can get it. It’s something that my colleagues at the Innovative Genome Institute and I are working hard to do.”
“One of the keys to all of this is delivery. We know that CRISPR works well, but the challenge is how to bring the molecules to the cells and tissues where they are needed. This is an area where we need innovation. We need smart people thinking creatively about how to do this effectively in specific types of cells. I feel very confident that this can be done. And this would be a way to reduce the costs and make this technology widely available worldwide.”
“Safety and efficacy are obviously very important; we want to be sure that CRISPR is being used in an ethical way as well (there is the report released from the National academies and addresses particularly the use of CRISPR in the human germline and putting in place international guidelines for responsible use in clinical settings).”
“Cell technology requires cells to be manipulated and is typically being done ex vivo, collecting, manipulating and then returning the cells to the patient. I imagine a day when we have not to do that, and we actually have a way to manipulate the cells in situ. Imagine sickle cell disease, for example, rather than having to do bone marrow transplantation for each patient, we have a way to deliver CRISPR to the right stem cells in the bone marrow for manipulation in the body effectively and safely. It would be absolutely transformative. This is why we need to make this technology available to all of those that need it.”
[About multigene diseases] “CRISPR is going to have an impact in uncovering the genetics of diseases and figuring out how genes interact in ways that are difficult to predict but you can now dissect through manipulation. That is going to take great work for sure. But having a technology that allows manipulation of specific genes is clearly key for addressing this kind of question.”
“And then suppose you knew a set of genes that need manipulation to affect a disease outcome, then the question is for the technology how to manipulate efficiently and safely a large number of genes. I’ve seen extraordinary data, especially on plants where a collection of genes can be manipulated at one to get the desired outcome. I think the technology is going to go there, it’s going to take additional clever work.”
“One of the things exciting about CRISPR is that it’s a very versatile platform. We understood that some CRISPR proteins have the ability to report on detection of an RNA or DNA sequence cutting a reporter molecule that has a fluorescent dye appended to it. This can be done for RNA detection using the family of enzymes known as Cas13 or for DNA detection using the family of enzymes known as Cas12.”
“There is a number of groups working on this in commercial and academic settings, and I think that is exciting to think at the potential of this to provide alternatives to PCR or other kinds of amplification-based technologies for coronavirus detection in particular. But this a programmable technology that can be easily harnessed to detect other viruses and to prepare for future viral challenges that I have no doubt we will have to deal with in the future. I don’t doubt we will see continued advance in the technology and new clever ways to use that fundamental mechanism to manipulate the genome.”
[Abouy CRISPR as antiviral treatment] “CRISPR is a great technology to understand Sars-Cov2 and its interaction with the host. But using it therapeutically will be hard, at least in the short term because of this delivery challenge and because we are still learning a lot about the biology of this virus and how it interacts with the immune system that seems to be fundamental for why people respond quite differently to the same virus.”
[About young researchers using CRISPR in the next 15 years or so] “You are the future and I’m so excited for the opportunity you have. It’s an extraordinary time to be in the biological sciences and there is so much to be done and my fervent hope is that CRISPR is a wonderful tool for all of you that allows you to ask questions that were impossible to answer previously. Science is about asking and answering questions and trying to develop new tools that allow us to ask the future set of questions.”