Nicola Patron is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an engineer than a biologist. Their focus at the lab is on plant-pathogen interactions, and her aim is to produce constructs and edit genomes so as to make plants, and agricultural crops in particular, resistant to disease. They also devise biosensors designed to elucidate the molecular interactions that go on between plants and their pathogens.
As Patron explains it, her work has always been focused on gene transfer, from transgenes to plants, chloroplast to the nucleus, or pathogens to their hosts. I spoke with her about what motivates her research, the MoClo Kit she and Sylvestre Marillonnet share with the scientific community via Addgene, the struggles of plant scientists and how they work to overcome them, and why she spends some of her time engaging with others on Twitter, among other things.
MoClo: Everybody's favorite parts for plant synthetic biology
Addgene: Tell me about the Golden Gate Modular Cloning (MoClo) Kit. What does it include?
Patron: Sainsbury has a mandate to share and educate and enable plant science in general. We are funded by a charity and since we are making all the tools for plant research here and validating them, it made the most sense to us to enable other people to do that too. We don’t have the capacity to send tools out to people ourselves, but we’d really like for people to use them.
The MoClo Plant Kit makes it really easy to clone genes for expression in plant cells. Sylvester and I chose the 96 most useful things for day-to-day life in a plant lab, so it includes everyone’s favorite promoters, plus a few new ones that are not used so much but have comparable expression levels to things people do use a lot. The problem in plant science is there is a paucity of resources. We wanted to expand the toolkit. We have included a wider range of terminators than are commonly used as we have tested them to show that they work well in plants and give similar stability to transcripts as the ones that people use all the time. Now there are many, so if you want to make a 10-gene construct, you don’t have to worry about repeating elements anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence can be challenging in plants because of the auto-fluorescence of chlorophyll, so we looked at newer fluorescent proteins that get around that problem.
MoClo is really a modern toolkit for those working in plants, including many things people already use. It makes the switch to Golden Gate Cloning easier because individual labs will not need to ‘domesticate’ all the basic and necessary elements in order to start using the MoClo system. The paper also gives quantitative data for regulatory elements, which has not been available for plants.
Addgene: Will the MoClo tools work in any plant?
Patron: The MoClo kit is entirely for dicots, not grasses and wheat, but we do work in wheat here. We do lots of work on wheat rusts and we are collecting sequences that will provide a similar resource for those involved in monocot engineering projects. The MoClo tools will work in any dicot, but they were developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is a tobacco relative and a good model for plant disease because it is susceptible to a lot of pathogens. It’s also easy to express genes in, which makes it a good laboratory workhorse.
Tips & tricks
Addgene: Do you have any particular tips for those looking to get started with MoClo?
Patron: The parts will be familiar to most plant scientists. It’s really a matter of choosing the parts that you need from the kit and getting started making your own constructs with Golden Gate MoClo. The barrier for those switching to Golden Gate is a conceptual barrier. Sometimes people don’t know how to start. They might want to use our promoters and terminators, but then they aren’t sure how to convert their own coding sequence to the MoClo system. It’s quite straightforward really and we’ll be producing an instructional video soon to make it easier for people to use the Golden Gate cloning system especially with plants in mind. When it’s ready, I’ll make those available on my website.
The thing is: it’s so easy once you are in the lab. There is no PCR, no gel purification; the whole cloning process is so simplistic once you have your parts. That was the motivation for releasing a large parts kit. Once you have so many parts available in your lab - if you get the 96-well kit - everything is so easy. We have undergrads come in for short projects and they end up making 30 constructs. They’ve never done any kind of cloning and they don’t have any prejudice about it. They just start and go ahead. I can see science speeding up.
Addgene: Are there other tools you are working on for plants?
Patron: Our lab and others at the Sainsbury are working on a lot of CRISPR tools for editing plant genomes. We will definitely plan to share those tools with the plant community. We’d like to add constructs to allow people to, for example, knock out a gene with simultaneous complementation, with a knockout and expression construct on the same piece of DNA. We do a lot of protein-protein interaction work and we’ve gotten some tools back from Addgene recently to explore their use in plants, including FRET tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools for plant science based around those technologies in the near future.
Science on Twitter
Addgene: You are active on twitter @nicolabiologist, where you tweet about plant synthetic biology, science communication, women in science, and many other topics. What’s the value of engaging about science in that way for you?
Patron: I started my Twitter account recently inspired by an academic in literature who gave a talk to the women’s research network here. She talked about the advantages of talking to people and representing yourself as a real person, so that students see academics as well-rounded human beings. It’s also a great communication tool for work. Most Sainsbury scientists are active on Twitter. Sometimes it’s a good way to find out what’s happening in the office next to you. There is an active Twitter scene of iGem teams that is really useful also.
There is also a U.K. organization called Athena SWAN that is a charter for advancing women in science. Universities sign up and in the first year there is an analysis of women in faculty, how they are doing. They look at structure, impediments, the representation of women on committees, and how the culture of the university impacts the number of women reaching senior positions. There has been a lot of engagement with the Athena SWAN Charter and it has really strengthened the network of female scientists. Twitter has been really good for that too. I’m more in touch with a remarkable number of female scientists whose tweets I find really inspirational. They share experiences, articles about women in science, and the sometimes stupid or uncomfortable things that have happened to them. To have that conversation out in the open is really important.
Addgene: Is there anything else you’d like to say to our readers?
Patron: Remember plants. Every time you look at your plate, think about how much that’s on it are plants, and the animals you eat ate plants too. Think about how many people are starving. I think plant biology is massively underfunded considering how important it is. I would really like to reach all those amazing students that want to have a big impact on the world and tell them to work in plant science. I think their chances of doing something amazing will be equally high as for those who go to work in cancer biology.
References:
A Golden Gate Modular Cloning Toolbox for Plants. Carola Engler, Mark Youles, Ramona Gruetzner, Tim-Martin Ehnert, Stefan Werner, Jonathan D. G. Jones, Nicola J. Patron, and Sylvestre Marillonnet. ACS Synth. Biol., Just Accepted Manuscript, DOI: 10.1021/sb4001504. Journal.
A modular cloning system for standardized assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed.
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Topics: Synthetic Biology, Plasmid Cloning, Plasmids
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