jimofoz

Hey did you guys get a positive result in the end for expressing GFP or insulin?

Hey I realize the contest is over so that you guys may not be checking these comments any more, but one other question(s) I had was - how will you separate the OMVs from the bacteria to make a topical formulation? And how will you ensure that the OMVs actually have a CAS9 endonuclease and and guide RNA in them? I saw this news (https://unews.utah.edu/pain-in-the-neck/) about researchers using CRISPR to epigentically turn down inflammation, and I have a mate who has really bad Eczema for which nothing offers any lasting relief, and was wondering how many other problems/gotchas there are before demonstrating this kind of thing in a mouse model of Eczema?

Well done. For use in humans, what about the LPS? How do you avoid this causing toxicity? Something like this? "Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins" https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4404585/

Any news on this project, or is it dead?

Any news on this project, or is it dead?

I am asking about antibody production, because currently this is done in very expensive bioreactors. maybe it is done in human cells? I am not sure. In addition the antibodies have to be administered sub cutaneously, like insulin, so developing oral antibodies protected be algae cell walls could be useful. You might be able to produce DARPins in the chloroplasts instead of antibodies, as they can be produced in E. Coli.

I am asking about antibody production, because currently this is done in very expensive bioreactors. maybe it is done in human cells? I am not sure. In addition the antibodies have to be administered sub cutaneously, like insulin, so developing oral antibodies protected be algae cell walls could be useful.

Are you guys aware of this Outer Membrane Vesicle project on experiment.com?: https://experiment.com/projects/crispr-capsules-packaging-cas9-with-bacterial-outer-membrane-vesicles

I'm assuming that if successful, this technique could also be useful for in vivo gene editing? What would be the salient challenges in using this for gene therapy/editing? The UK Cystic Fibrosis Gene Therapy Consortium is developing a Lentivirus for inserting a correct gene into cells lining the lungs, could your technology be useful? Do OMVs nebulize well?

So are you actually delivering the full CAS9 protein to bacteria in these outer membrane vesicles, rather than just a plasmid to express the CAS9 protein inside the bacterial cells? Has anyone else tried this before? In vivo, wouldn't the animal's immune system attack the OMVs before they can reach their target bacterial cells?

I was thinking more about using the chloroplasts ability to survive the stomach and deliver proteins into the intestine. Growing the bacteria inside the algae would be hard. But how about adding them to the algae just before ingestion? I know that in various human diseases where a genetic error results in a person being deficient in a lysosomal enzyme, these enzmyes can be packaged/taged so that upon injection they are traffic to the cell lysosome. I was guessing that you might be able to do the same thing with these beneficial bacteria to get them into the gut orally rather than via a fecal matter transplant.

Also Todd Rider is trying to crowdfund development of a treatment called Draco that could deal with all double stranded RNA viral infections. Could you guys produce this molecule in chloroplasts that could be ingested by fish? This could make the value proposition of your company even stronger. https://www.indiegogo.com/projects/dracos-may-be-an-effective-cure-for-viral-diseases#/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3144912/