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Create GLOWING PLANTS using synthetic biology and Genome Compiler's software - the first step in creating sustainable natural lighting
Create GLOWING PLANTS using synthetic biology and Genome Compiler's software - the first step in creating sustainable natural lighting
Create GLOWING PLANTS using synthetic biology and Genome Compiler's software - the first step in creating sustainable natural lighting
8,433 backers pledged $484,013 to help bring this project to life.

Glowing Plant March update

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Hello backers,

Here's March's update:

Summary

  • We are still working to debug the latest issues with the Lux plants. 
  • We have fixed the challenges with the Fluc v2 plants. 
  • We are expanding the maker kit beta tester program so anyone can join
  • We are still planning for a non-accredited investor fundraise launching around May 16th

Autoluminescent plant Lux v1

We are still in the process of regenerating the plant. The first batch of regenerants didn't glow stably, although we did find one weird plant which was glowing a little but which then stopped glowing which was a very strange and unexplained result. 

We've updated our hypotheses for what is going on based on the results from the last month, in particularly the lack of success from any of the regenerated plants. These are the three most likely reasons for the challenges we are facing:

1. The most likely scenario is premature removal of selection pressure leading to false positives and seedlings that do not include the six lux genes. In order to speed up the regeneration process and shorten the time to getting seeds we removed the selection pressure after 2 month. We did this because the plants grow much bigger much faster once selection pressure is removed. It seems to be working but the academic literature suggests keeping the plants under selection pressure till the roots have formed so this may have been a mistake. Now we are keeping the plants longer under selection and this has reduced the number of regenerants dramatically, which supports the hypothesis. This longer selection does lead to slower growth delaying our progress and means we don't have clear results yet. We are waiting for the next batch of plants to root before running the assay to see how well they glow.

2. The second possible issue is caused by the low transformation efficiency of co-bombardment with a large construct: We have chosen to do co-bombardments (lux and marker on constructs) to ease the marker removal step (as we don't want to spread herbicide resistant genes into the environment). Under this hypothesis, only a portion of the selected regenerants would have received the lux construct reducing the efficiency. With our Fluc plants that proportion was 50%, but with the larger Lux construct  we expect that proportion to fall even lower. Starting last week, we have been testing using a single construct for bombardment, which means almost all of the regenerants selected should glow. If this works we will have an additional challenging step to remove the selection marker as we won't be able to out-cross the marker as it will be co-inserted with the glowing genes. We plan to explore the CRISPR/Cas9 system to achieve this if we have to.

3. The final possibility is gene silencing of the construct. It's possible that the anomalous plant which glowed briefly for a few days is evidence to support gene silencing. The promoters we are using are also used by the plant for essential life functions, so silencing should kill the plant (in theory) and hence this shouldn't happen. Gene silencing is poorly understood though and not many groups have created transgenic plants with this many genes inserted so there's plenty of uncertainty here. Generating a large number of independent transformation events is one way to address silencing issues as the problem may be more or less sever depending where in the genome the construct is integrated. In the meantime, we have also performed a floral dip to transform Arabidopsis with the lux construct to test if silencing is happening there (this is a much faster regeneration process).

Fluc v2 plants

We discussed the germination problems in the last update.Our hypothesis about anther sterility was correct and now we have nicely germinated T2 lines growing in our growth tent. These plants will be tested for luminosity and marker presence next week. Once we find one that glows nicely without a marker sequence, we will be ready to grow that one to make it's own seeds and ship out like the Fluc v1 plants - so this is back on track.

Fluc v2 plants with fixed anther problems
Fluc v2 plants with fixed anther problems

Maker kits

We shipped the first maker kit to a school this week which is exciting, looking forward to hearing about the kids reaction. Our revised permit guidance document also seems to be working better and one beta tester got the permit approved within a month which is awesome. The kit is still in beta, but we are now ready to roll out to anyone who wants to be a beta tester. If you backed the maker kit and would like to be a beta tester (and aren't already part of the program) then please get in touch with us.

Title III fundraise

We are still planning a public equity crowdfunding campaign. This is provisionally scheduled to launch on May 16th. More on that in a future update.

All the best,

the glowing plant team.

OpenROV, John Floresca, and 19 more people like this update.

Comments

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    1. Antony Evans Creator on

      Tyler: yes, we have a problem with the terminator in the moss strain and we are having to rebuild it.

    2. Missing avatar

      deleted on

      This user's account has been deleted.

    3. Antony Evans Creator on

      Brian: we want the minimum investment to be as small as possible to enable as many people to invest as possible, not just already wealthy. We still need sign off from lawyers as this is a new thing but hopefully the minimum will be $100

    4. Antony Evans Creator on

      John: email us and we'll get you started with the maker kit

    5. Antony Evans Creator on

      John: send us an email and we'll get you started on the maker kit

    6. Brian Roush on

      Antony, any word on what the minimum investment will be for your funding round?

    7. Antony Evans Creator on

      Sean/Six: Thank you for the kind words, your support and continued encouragement means a great deal to the team.

    8. Six on

      "this is the most satisfying Kickstarter I've backed personally" +1 @Sean Houlihane - Of the 40 something projects I've backed and the thousands I've researched this project has consistently provided through, transparent updates and demonstrable progress.

      I'm easy to find online and am very vocal about the blatant deception present on many KS projects, this is not one. This project may be late but it is awesome to watch and be part of. Remarkable work.

    9. Marie McHugh on

      Thank you, Great Job! Keep Working!

    10. Antony Evans Creator on

      Tomas: Thanks for the interesting suggestion. At this stage we aren't even sure silencing is happening so this kind of analysis is a bit premature.

    11. Sean Houlihane on

      I know this is nowhere near the original plan, and maybe quite a few backers are disappointed with the progress, but this is the most satisfying Kickstarter I've backed personally. Most of the physical rewards I got from other projects are unused in a drawer (assuming they arrived), and I don't feel I contributed much to establishing a market for the products - but here you have real interesting things going on (even if I'm only guessing about what some of it means)

    12. Tomas on

      "The promoters we are using are also used by the plant for essential life functions, so silencing should kill the plant (in theory) and hence this shouldn't happen. "

      Have you thought about looking at it with FISH? See if it's in a different spot than where the promoter locus should be? As in, it might be silenced by having being 'moved' to areas bordering the INM and silenced conformationally.

      You could get a killer paper out of it, answering the question of whether genes lie close to the INM because they are inactive, or does lying close to the INM make genes inactive.

    13. Antony Evans Creator on

      John: Please send me a private message or email to join the maker kit beta, we can't track the comments easily.

    14. Antony Evans Creator on

      Thanks for the support backers.

    15. Lea B. Sims on

      Can't tell you how much our family loves your updates. My kids backed this project with saved up allowance money and they are super proud of all of the work you are doing. They love to talk all about it with family members and anybody who will listen. :)
      Thank you for all that you are doing!
      Your biggest fans,
      The Sims

    16. Missing avatar

      Marcia Morrison on

      Fascinating! This is my favorite part of backing Kickstarter projects--I get to hear about all sorts of cool stuff, and learn new things too. Keep up the good work!

    17. Paul Cline on

      Thanks for the update.

    18. David MacDonald on

      I basically understand almost none of this science, but I do love reading these updates! Thanks for taking some time away from the actual research to keep us informed.

    19. Missing avatar

      John Floresca on

      How can I be a beta tester or get a makers kit?