Glowing Plant September backer update
In this months update we've got some information on the progress with the autoluminescent glowing plant, some information on the start of our attempts to make glowing moss, an educational video explaining homologous recombination, update on the fragrant moss, and a promo code for synbiobeta where we will making our first public demonstration of the fragrant moss.
In continuation of the effort, we are still in the process of screening and regenerating tobacco tissue bombarded with our full construct. We are still lacking a plant with all six genes but now have many lines with three to five genes. As we explained in our last update, we are going to compliment these lines with a line that contains the missing genes. All of our lines are missing luxC thus, we started work on creating a transgenic tobacco line by inserting a DNA piece containing the luxC cassette. Bombardments have been initiated and screening and regeneration of these lines are in progress.
Plants derived from this screen will be crossed with the previous lines to complement the missing genes. This seems like an extra step, however, remember we are using a new selectable marker cassette which does not need to be crossed out: Overall, the number of generations the plants have to go through to get the final shipable seeds remain the same.
Probably, you all are aware of our Fragrant Moss and that we've started work on a Glowing Moss in case the tobacco plant doesn't work out as we want it to. Our first attempt is a basic demonstration using the Firefly luciferase cassette, as we did previously in tobocco. In parallel to the proof of principle, we are working on creating different versions of constructs containing the six autoluminescence genes. Some of you might wonder why we should make new constructs when we have the working one for plants. There are couple of reasons:
- First of all, the dicot promoters do not perform well in moss. For our autoluminescent plant, we are using endogenous promoters from Arabidopsis. However, researchers have shown that the performance of these promoters are minimal in moss. In order to achieve a strong constitutive expression of the pathway, we need to use strong promoters from monocots such as corn or rice that are known to perform well in moss.
- Another reason is based on the fact that moss transformation uses a different mechanism from plants. While we force the insertion of exogenous DNA into plants using biolistic methods, moss uses a system called homologous recombination. Detailed explanation by James can be found in this video below. As we only have a couple of known strong promoters in moss, the way to create a six gene construct requires repetitive use of these promoters. Currently, we are not sure how the repetitive elements will affect the transformation process. In order to overcome any difficulties the repeats provide, we are in the process of designing various ways of introducing the DNA pieces into the moss genome. More information will follow in future updates.
What is homologous recombination?
One of the unique features of the strain of moss we are engineering is that it can do something called homologous recombination. This is a specific mechanism for how DNA interacts within the cell, typically found in prokaryotic cells rather than eukaryotic cells, which is explained by James in this video (sorry for sound quality, use the subtitles if you can't hear properly):
Update on fragrant moss
We are ramping up production of the Hygromycin resistance-free fragrant moss line which is the version we plan to sell and distribute to keep funding the glowing plant research. After some initial problems with a fungus infection in August we've cracked the process for scaling up and manufacturing larger amounts of the moss. Currently we can double or triple the amount of moss we have every two weeks.
We are also testing various growth conditions for moss that will enable it to grow faster, and experimenting with ways to track growth of the moss on agar plates. For instance, we plan to buy a tissue homogenizer that will allow us to more efficiently break apart clumps of moss to spread on several different plates, enabling faster propagation and a shorter doubling time.
Looking forwards, we have a list of ten candidate scents to engineer in moss in the future. We will also be starting to sell and distribute the moss once we have enough to ship in volume. We are still finalizing the plan for exactly how we are going to do that.
Synbiobeta + demo of Fragrant Moss
We are excited to announce that we will be demoing the fragrant moss at the Synbiobeta conference in San Francisco on October 4th and 5th. It was at Synbiobeta last year that we met our Danish collaborators on the moss, so it's appropriate that it's the first location for a public demo. Glowing Plant backers can receive 20% off the registration price if you book before the end of September: just use this link: synbiobeta.com/conferences/san-francisco-2016/ and promo code SBBPARTNER20.
Hope to meet you at Synbiobeta, otherwise we'll be in touch with the next update.
The Glowing Plant team