Re: [DIYbio] Re: Dwarf tobacco?

My humblest apologies Andrew! I think the email was either snagged in my spam net or I simply didn't get a chance to write back. The poster was me. I use my Gmail account to post to DIY bio so I can keep tabs on what's important and interesting (DIY Bio) versus the crap ton of biotech supplier ads and the occasional business email that floods my NYB inbox. Truly sorry that I didn't get a chance to write back!

I don't want to make my work seem like something its not. I work from my home lab soon to be pro lab in a separate location. I started a company because I had a few ideas I wanted to work on while trying to generate some income for the lab. I began with orchid micropropagation and sold to a few hotel chains in the local NYC area but that all stopped when I fell in love with plant genetic engineering. I decided to turn the company into a non-income generating R&D model until I do manage to make something worth selling. Im almost entirely self taught aside from an almost finished BS from Stony Brook University so I cant really advise on whats best from an academic stand point. Im doing this all on my own which is terrifying since if I mess up its all on me but liberating since I don't have to be pressured by a PI or grant deadline etc. Its a hobby that turned semi-pro…nothing special.

As far as agrobacterium and my knowledge of it is concerned, strains like GV3101 EHA105 etc are up for deregulation but until then its treated as a plant pathogen and thus BSL1-2 grey area. In some cases under specific conditions it can and will transfect mammalian cells. Moving plant pathogens or material carrying such across state lines in the USA is a felony. I have absolutely no clue how on earth Glowing Plant Project people got around this…or did they disclaim everything in superfine print? Wild type agrobacterium transfers oncogenes that produce similar cytokine compounds that trigger cell proliferation (cancer). The hormones they code for (genes IaaM, IaaH, and IPT from the Ti plasmid) are also considered carcinogens but in large concentrations. Until the proper authorities concluded beyond any doubt that the disarmed strains are in fact disarmed and can in no way transfer their genes to other things (which would defeat the purpose of agro) they will keep the bacteria regulated to a degree.

As for my “research”, I have nothing formal published yet but expect a small article to be done soon. My focus is on using polyethylene glycol as a means of transforming plants and more importantly the plastid (chloroplasts). Gene guns are expensive to buy and expensive to shoot. The DIY derivatives are unreliable at best and known pressures for plastid transformation is in excess of 1000 PSI so that kind of pressure I would avoid. Its also very inefficient. About 5% transformants in comparison to the 80%+ with agro but it can only target the nucleus. The use of alternative carrier nanoparticles like tungsten tend to be slightly to severely bioavailable or bioreactive which may induce defensive apoptosis. Either way in my opinion the gene gun and agro are not the answer but a convenient solution to genetic engineering in higher plants. I like PEG and protoplasts but they are tough to handle. Im working on making the process as fool proof as possible. Worst case Ill try to publish it in PLoS ONE or just a detailed entry on openwetware.org.

I'm trying to organize my life so that I can have time to post to my blog which is or will be 100% plant biotech. For now I post religiously on Facebook so you can friend me if you’d like and follow my rants, anecdotes, and cooking-show-like protocols.

Sent from Windows Mail

From: Andrew Willoughby
Sent: Tuesday, February 4, 2014 12:18 PM
To: diybio

Well here's a copy of the email I sent to NY Botanic's email address:

I stumbled upon your website from a poster on the DIYbio google group/mailing list. Sorry if I try to take this opportunity to mine you for questions; plant biotech is my dream. 

Are public plant biotech labs common? Google searches don't seem to turn up much.

Is there anyway I can browse your research? (Papers that is)

I've applied to a disgusting amount of schools, 7 with plant science/ biotech/ bioengineering -ish programs:

University of Illinois @ Urbana-Champaign: Bioengineering and Horticulture
Colorado School of Mines: Bio E

Cornell: Plant Science

CalTech: BioTech

University of Southern California: Biotech, with a botany research focus

Colorado State University: Horticulture

University of Oklahoma: Botany

Are any of these schools widely recognized in this field? It's hard to find any kind of reliable source analyzing a school's academic quality in botany. I guess not enough people are interested, sadly.

If getting a job in a lab like NY Botanics, what other jobs are out there for plant scientists/ plant biotech-people? I'm strongly considering double majoring in Chemical Engineering as well to be more hireable.

Is there any other advice anyone out there can give a stressed out 18 year old trying to decide, if not the rest of his life, then the what opens the door to it?

So are you working on making gene guns more efficient? If I remember correctly from watching the Glowing Plant project's videos, you don't need the agrobacterium if the gene gun works.
Is agrobacterium regulated because of horizontal gene transfer-phobia?

I'm sure I'll think of a few more questions
Andrew Willoughby

On Feb 4, 2014 10:49 AM, "Sebastian Cocioba" <scocioba@gmail.com> wrote:

I have a personal lab at home in NYC and will soon be moving it to a
business address. I've built my lab specifically for plant biotech and
all was procured through eBay and university auctions. Its amazing how
well set up you can get with a little patience and some negotiating. I
don't think I've spent more than $4k in total but have the value of a
few tens of thousands.

Like I've mentioned previously, the limiting factor in doing plant
biotech DIY or otherwise is agrobacterium regulations. Im working on a
way to eliminate the need for bacteria in plant transformations all
together and make it available as a kit or open source protocol using
commercially available ingredients. This just put even more fire under
my ass to get this to work.

If you have any questions just ask. I'd love to help make your research
dream a little more tangible. Its super rare to find people that are
interested so by all means ask away! :)

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D From: Andrew Willoughby
Sent: 2/4/2014 9:22 AM
To: diybio@googlegroups.com
Subject: RE: [DIYbio] Re: Dwarf tobacco?
Where are you doing this? I'm a high school senior and it's my dream
to do plant biotech. I'm trying to make college fit that.

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