Making oral vaccines and insulin in microalgae

[antomicblitz]

Hey everyone!

I wanted to take this opportunity to share a cool project that I think could be really exciting and I want to get the community involved.

Making Inexpensive Insulin and Oral Vaccines Using Microalgae

I am working with a couple of people from UC Berkeley, UC Santa Cruz, Ireland, and New York to try and run a pilot study demonstrating the feasibility of using microalgae to produce therapeutic compounds like oral vaccines and insulin. By using insulin for our pilot study, this also allows us to collaborate with the Open Insulin Project since the genetic machinery in both E. coli and the chloroplasts of microalgae are bacterial in origin and consequently are very similar. We launched our experiment page a couple of days ago and we could really use some help getting the word out. If you want to be directly involved, please send me a message :). Please help spread the word by liking or sharing on social media, and if you have some extra dough, consider putting it to a good cause.

P.S. For those who know Sebastian Cocioba, he's also donating his amazing expertise (and hopefully some of his chloroplast transformation vectors if we can get some funding) :)

Warmest Wishes,

Antonio Lamb

[BraveScience]

Hi Antonio,

Cool project.Nice to see somebody else working with algae, especially given the topic.

I may have some extra dough, even though is cyanobacteria based (metabolic engineering and synthethic biology)

I have some questions for you, hope you can answer :)

1) do you plan intracellular expression or extrusion into medium of insulin? 

2) are microalgae suitable (i.e. genetic toolbox for protein expression and/or extrusion)? (plastidial expression may add layers of complexity to your system as well)

have you considered cyanobacteria for the study?

Admirable project, wish you all the best!

Cheers,

Fede

[antomicblitz]

Hi Fede,

Thank you :).

So to answer your questions 

We are planning to do intracellular expression strictly within the chloroplast. We are actually doing this to avoid many of the issues that occur because of nuclear expression, and it actually helps reduce the layers of complexity. I'll explain why.

There are a lot of advantages to expressing genes in the chloroplast because of the bacterial-derived expression machinery and the membrane bound structure of the chloroplast. 

1. Since the chloroplast is inherited maternally, there is a low chance of a foreign gene escaping in nature,

2. Since there are multiple chloroplast copies in the cell, there is a higher level of expression

3. Unlike expression from the nucleus, gene silencing doesn't seem to happen in the chloroplast

4. The chloroplast has enzymes that are capable of facilitating disulfide bond formation in proteins. This allows a wider variety of proteins to be expressed stably. 

5. Foreign proteins that would normally be toxic to the cell can be encapsulated within the chloroplast membrane

6. Any bacterial genome can be inserted into a chloroplast genome since it is of bacterial origin itself.

7. The cell wall and chloroplast membrane help facilitate delivery of oral vaccines. The cell wall allows a protein to survive the digestive track and allows the protein to be absorbed through the stomach. This is ideal for oral vaccines.

I encourage you to study the scientific literature here, here, and here if you want to know more.

Our reason for using insulin is because there is a lot of literature to guide the experiment, so we know it's been done before. Also, if oral delivery of insulin ever becomes possible, algae might be a good way to do it. you never know.

I actually have not considered cyanobacteria, but given that they are very similar to chloroplast, they would probably be great test organisms as well. You brought up something that might be really helpful that I had not thought of. Thank you :). 

I hope this has answered your questions. Please let me know if there is anything else you'd like to know.

Antonio

[Ravasz]

Hi,

This is a very cool project, kudos to you guys.

I have been developing a similar pipeline myself, but I started with the growing part and building automated bioreactors first to grow the algae in before I start with any transfections. If you think I can help you in any way just let me know, it'd be great to team up with like-minded people.

My first thought when looking at your project was the same as Fede's: I would imagine using cyanobacteria would make your life a lot easier. That's what I am planning anyway...

My second was that insulin will just get digested if eaten with algae, and purifying it from chloroplasts would probably be more expensive than the current production methods. Also, oral vaccines tend to be entire virus capsids or even bacteria spores not single proteins that are expressed alone - although I admit I don't know the topic really well, so I am happy for any education on this. My idea was to express GFP first as a proof-of-concept, and then try to build some flavour compounds (although I haven't actually chosen any as they too seem rather difficult).

Either way, if you are interested in further discussion, I'd be very to talk to you guys about more details.

Cheers,
Mate

[antomicblitz]

We would love to have you on board, my friend :). And to answer your question, the utility of orally administered vaccines and insulin expressed in lettuce and tobacco chloroplasts has already demonstrated some promising results with the prevention of insilitis in diabetic mice.

http://www.ncbi.nlm.nih.gov/pubmed/17490448

[John Griessen]

On 12/07/2015 04:34 PM, Ravasz wrote:
> I started with the growing part and building automated bioreactors first to grow the algae in before I start with any transfections.

What are those like? Do you plan a continuous flow system, or batches?

[antomicblitz]

Again, if anyone is interested and would like to collaborate, contribute their expertise, bounce ideas, or provide materials (such as cyanobacteria or algae strains, bioreactor design tips, etc.), let me know.

My email is amlamb(at)ucsc.edu. I respond pretty promptly.

Antonio