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Offlinechibiabos
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Registered: 03/16/17
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Re: CRISPR/Cas9 [Re: FishLevelMidnight]
    #24742450 - 10/27/17 10:59 PM (6 years, 3 months ago)

Quote:

fishermansjc said:
This is definitely true, but you can have variation making CRISPR animals. If you start from a wildtype population and use CRISPR to induce mutagenesis than you will still have all the diversity of the wild type population (and have variance in your genetics) EXCEPT at the intended CRISPR target site.


In theory you can also use CRISPR to introduce naturally occurring alleles back into a population that has undergone a bottleneck or whatever.


And yes, we cannot anticipate everything, but becsuse we understand the mechanism that CRISPR acts on, we know where and what to look for in terms of off target effects.
To say we don't know where or what to look for is like saying that you plan to change one of the components of your bulk sub and then someone asks if that caused the paint in your grow room to change, some things are just not going to happen when you add or delete genetic code



I think that they mean more in the context of systems biology than whether you can walk away from an experiment and make a valid argument that your observations are a result of your CRISPR protocol.  :shrug:


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OfflineShabbyRabbit
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Re: CRISPR/Cas9 [Re: chibiabos]
    #24742507 - 10/28/17 12:08 AM (6 years, 3 months ago)

This is an off-hand comment.

So while thinking about how to use this tek to make mushrooms taste better I stumbled upon this article "How CRISPR is changing the food industry"

Thought I'd share.

Edit: Think strawberry mushes...mmmmmm.  Also, here is an interesting site with many manuals for the kits they sell.  Very informative.


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Edited by ShabbyRabbit (10/28/17 12:19 AM)


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OfflineWiiiiilson
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Re: CRISPR/Cas9 [Re: ShabbyRabbit]
    #24744659 - 10/29/17 02:14 AM (6 years, 3 months ago)

Cool article!

It mentioned virus resistant pigs. Treating viruses is one use of CRISPR that doesn't require editing of the genome, but rather directly targeting the viruses themselves.

Many viruses are extremely common (like Human herpes virus 6, which almost everybody has) and which sporadically reactivate, typically when someone is already stressed or weakened - sometimes with terrible consequences.

Lots of these latent viruses have been linked directly with the development of certain cancers and autoimmune diseases and many are members of the herpes virus family (HHV4 = lymphoma, SLE, MS. HHV6 = lymphoma, sarcoma, cervical cancer and brain tumours. HHV8 = sarcoma, lymphoma). Another major target - HIV. It might be more difficult to target a virus that can change so quickly, but it has to be better than trying to inhibit it with drugs.

These viruses could be eradicated from the population one day!


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OfflineChaostoOrder
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Re: CRISPR/Cas9 [Re: Mike O Voidenski]
    #24744680 - 10/29/17 02:41 AM (6 years, 3 months ago)

Quote:

Mike O Voidenski said:
Here is a pdf link to a step by step guide on how to perform successful CRISPR experiments.
http://www.synthego.com/wp-content/uploads/2016/12/How-To-Perform-Successful-CRISPR-Experiments.pdf?hsCtaTracking=d846003c-6136-425e-ab48-83cf0273bd75%7Cab96c0a9-4a8c-4b70-8a63-9bae54639f25

I edited the intro post to include this link to help grab the interest of thread skimmers.

I read that RR was able to improve PE by crossing monocaryon myc of PE and another better spore producing mushroom to come out with PE6. Maybe someone can now cross a woodlover with an easier to grow species. I know some woodlovers are very closely genetically related to some more popular and easy to cultivate species. Another thought is now people can begin working on some mushroom super strains. Maybe even completely new species that are able to do new things.

Maybe with CRISPR being used to develop new mushrooms we will start seeing fungi capable of producing all kinds of new experiences and even curing all sorts of diseases that we were able to do little about before this new technology.

It appears that someone has already used CRISPR to improve a mushroom. I have the link edited into the intro post above.

Anything is possible with this stuff and weather you are ok w it or not people are experimenting with this stuff right now. I may go down this road eventually but I need to get more hands on mycological experience before I start playing mad scientist. It's probably a job for some more experienced mycologists..... Hint hint.




So I'm thinking the same thing. My theory is that this might actually be a good study on the grounds of plant evolution; that includes the fungi kingdom.  I'm thinking it would be a great study because we can maybe cross bread some of these species as normal evolution may have done had it not been for "over population" and "global warming". If there was a natural set course maybe some of these newly crossed strains would be the full result? IDK just my take on things, maybe I'm thinking way out there, if I am just tell me to think outside the glovebox hahaha lol


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Offlinesimmer
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Re: CRISPR/Cas9 [Re: ChaostoOrder]
    #24749382 - 10/31/17 08:27 AM (6 years, 3 months ago)

Not sure if this paper has been discussed here before but seems relevant to this thread: Enzymatic Synthesis of Psilocybin and a magazine article about it ‘Magic mushroom’ enzyme mystery solved

Researchers sequenced genes of cubensis and azurescens and identified genes for 4 enzymes used in psilocybin production, and some of the testing involved engineering E Coli to produce some of those enzymes.

I think that'll be the ultimate effect of bioengineering on psilocybin production - not engineering mushrooms to be better, but engineering bacteria to produce psilocybin.


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InvisiblebodhisattaMDiscordReddit
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Re: CRISPR/Cas9 [Re: simmer] * 1
    #24749392 - 10/31/17 08:35 AM (6 years, 3 months ago)

Was big news a few months back. Still above the pedigree of at home science for now


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OfflineFishLevelMidnight
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Re: CRISPR/Cas9 [Re: simmer] * 1
    #24749852 - 10/31/17 12:05 PM (6 years, 3 months ago)

If I had more knowledge on protein extraction, I would clone these enzymes from my P. cubensis into bacteria with like a His tag, then purify the proteins. With the right buffer solution you could simply add the enzymes in a tube with some starting material and produce actives.

It would be super cool for creating some of the other amines like using DMT as a start (vs using DMT in like the substrate/spawn; using it in a controlled biosynthetic pathway seems like it would have better yields).

Maybe I can simply clone the whole pathway into a bacteria and feed the bacteria culture with the starting product, then lyse cells to extract the actives (maybe with 190 proof EtOH, would be like doing it on fruitbodies but with a much higher ratio of actives to proteins/lipids).

I should switch my PhD topic! haha


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OfflineSemperViva
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Re: CRISPR/Cas9 [Re: FishLevelMidnight]
    #24750442 - 10/31/17 06:11 PM (6 years, 3 months ago)

Quote:

fishermansjc said:
If I had more knowledge on protein extraction, I would clone these enzymes from my P. cubensis into bacteria with like a His tag, then purify the proteins. With the right buffer solution you could simply add the enzymes in a tube with some starting material and produce actives.

It would be super cool for creating some of the other amines like using DMT as a start (vs using DMT in like the substrate/spawn; using it in a controlled biosynthetic pathway seems like it would have better yields).

Maybe I can simply clone the whole pathway into a bacteria and feed the bacteria culture with the starting product, then lyse cells to extract the actives (maybe with 190 proof EtOH, would be like doing it on fruitbodies but with a much higher ratio of actives to proteins/lipids).

I should switch my PhD topic! haha




How would you then go about separating the actives from the buffer solution without denaturing the enzymes?


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OfflineFishLevelMidnight
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Re: CRISPR/Cas9 [Re: SemperViva]
    #24750456 - 10/31/17 06:18 PM (6 years, 3 months ago)

"How would you then go about separating the actives from the buffer solution without denaturing the enzymes?"

Do you mean in the scenario I proposed in the first paragraph?

In which case, you could probably do some sort of extraction, maybe even centrifuge it (to pull down the proteins).

Worst case scenario you also eat some enzymes and buffer? :shrug:
We eat enzymes all the time so there is no issue there, and buffers bring the solution to levels optimal for the enzyme in question, so they are physiologically normal. Just like drinking a little saline.

Considering the active dose of psilocybin is ~20 mg, it wouldn't be all that terrible to consume a mL of buffer...


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Invisibleellomello
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Re: CRISPR/Cas9 [Re: FishLevelMidnight] * 1
    #24750565 - 10/31/17 07:11 PM (6 years, 3 months ago)

Mmmm buffer  :homerdrool:


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Offlinekrypto2000
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Re: CRISPR/Cas9 [Re: SemperViva]
    #24754136 - 11/02/17 10:15 AM (6 years, 3 months ago)

Quote:

SemperViva said:
Quote:

fishermansjc said:
If I had more knowledge on protein extraction, I would clone these enzymes from my P. cubensis into bacteria with like a His tag, then purify the proteins. With the right buffer solution you could simply add the enzymes in a tube with some starting material and produce actives.

It would be super cool for creating some of the other amines like using DMT as a start (vs using DMT in like the substrate/spawn; using it in a controlled biosynthetic pathway seems like it would have better yields).

Maybe I can simply clone the whole pathway into a bacteria and feed the bacteria culture with the starting product, then lyse cells to extract the actives (maybe with 190 proof EtOH, would be like doing it on fruitbodies but with a much higher ratio of actives to proteins/lipids).

I should switch my PhD topic! haha




How would you then go about separating the actives from the buffer solution without denaturing the enzymes?




After the enzymes have done their job it doesn't matter if they are denatured, they've served their purpose so you just have to focus on extraction and isolation.


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Offlineghaleon
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Re: CRISPR/Cas9 [Re: krypto2000]
    #24754720 - 11/02/17 02:41 PM (6 years, 3 months ago)

Would you be able to make glow in the dark cubes?


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