Experimenting with CRISPR Cas-9 at Home

[@zahratabatabaei5849]

Wow, I'm a biologist but the thing I was shocked with was the fact that you're experimenting at home while others may not be that interested to even study! Thanks for sharing!!

[@amymli]

Thank you for all that you do!

[@rania_hashim]

okay dude i love how you showed all your mistakes and was really genuine in the whole process

[@kothejunglist]

She made the mistakes so that we don't. :)

[@HugoSanchez-qq2oh]

Great video! I learned alot.

[@nephronpie8961]

@@mcruff3 I'm sorry, I was in a bad mental state at the time. I don't think anything can justify what I wrote back then.

[@mcruff3]

@@nephronpie8961 I hope you're doing better.

[@nephronpie8961]

@@mcruff3 thank you for asking. Yes, I'm doing relatively better now. Regaining the mental state of pre-covid times is turning out to be difficult, but I'm trying to take it slowly and learning more about myself in the process.

[@dawnsvale]

You could try mixing the Cas9 protein and the sgRNA first. Incubate for 20min at 37degrees celcius. This would allow the sgRNA to form a complex with the Cas9 (look up the structure of this ribonucleoprotein). Then, add the above complex, DNA template to your cells. After transformation, let the bacteria grow in LB broth without antibiotic first so they can recover from that heat shock and it will take some time for the DNA to be edited by the Cas9 complex. 1hr at 37deg would suffice (on a shaker if possible or shake every few mins). All the best!

[@amymli]

Woah! thanks so much for these detailed instructions I'll definitely keep them in mind the next time I do a transformation

[@rafalczyzyk9939]

@@amymli I support the above, but additionally you should include control plates. Before the CRISPR, include one plate with strep to make sure you cells do not grow on this and after CRISPR include one plate without any strep to make sure your cells actually survived the whole process (edited or not). Hence if you have problems, you have the means to discover where it went bad. Otherwise is cool what you do! Keep up!

[@Kuznecoov]

Cool. There is so small number of videos about crispr cas at home. Thank you for sharing 🐱

[@JasonWhittle1]

Looking forward to the update video. I'm really keen to see people doing this. It's insanely interesting.

[@stephenlangsl67]

Thank you for making and uploading this instructional video Amy Li. I just got through sharing it on Twitter.

[@amymli]

Thank you for the support!

[@rahulyaji]

Great video. Those explanations using paper cuttings were fantastic!

[@amymli]

Glad you thought so! I find that many complex topics in biology are best explained visually & temporally

[@jessykhalife5532]

Just watched your video with my gr 12 bio students! thanks for showing us that science can be done at home and double thanks for showing us that mistakes are real! Keep going!

[@amymli]

wow this means so much, thank you! if you're interested, you should definitely check out www.the-odin.com/ for more affordable genetic engineering experiments

[@rania_hashim]

AMY THIS IS SO COOL AND PROFESSIONAL OMG EVERYONE I WAS HERE BEFORE AMY BLEW UP

[@amymli]

lmaoo thank you!!! and yes rania was #first

[@IQTheRapper]

This was incredibly interesting. It's amazing how far biosynthetic research has come. The implications do trouble me, though. I hope someone doesn't ruin this for everyone, lol.

[@dawsonsmith3797]

[De-Extinction has entered the chat.]

[@Lima547]

Keep posting! Great experiment! I was born with PKD, a genetic disease, and 4 weeks ago I had my kidneys removed. So, for me, science is the only way I survive today (dialysis) and that will enable me to survive tomorrow and keep loving my family!

[@maziastro9932]

You earned a sub literally the best work I have seen and realllyyyyyyyy nicely presented and made things simple

[@remlapgarage]

This was so so informative, thank you and best of luck next time, would be great to see you do it again and succeed :)

[@amymli]

Thank you so much!

[@patrickwilliams997]

Failure is the mother of knowledge, keep it up! You're doing great!

[@abhaypal5155]

Nice! I gotta say your streaking technique is something new to me.

[@preli420]

That was an awesome video. Extremely fascinating.

[@amb-b1v]

Thank you very much for your video! I am from Russia, I poorly understood this topic and (oh my God) we were given a report on it!!! In your video everything is simply and clearly stated, every step and process. Thank you! You also nicely explained on drawings, ribbons and a bracelet! Спасибо, от души! ОАОАОАООАОАО великолепно!

[@amymli]

I'm so glad! Good luck with your report :)

[@sudegurdogan]

I am a molecular biology and genetics student, and this is very helpful for writing the lab protocol/process. Thanks for sharing ❤

[@peterxyz3541]

I came for “mad scientist at home”, I’m staying & subbing for the sound effects when pouring agar into dishes. 😂😂😂

[@dorfinhorg9458]

This is awesome! Just a heads up, when picking bacteria from a plate, you most always want to only pick one defined colony. That way, you know that all the cells in your growth are genetically homologous. Different colonies can have different genotypes.

[@amymli]

Thank you for the tip and explaining it so well! I'll be sure to keep that in mind for future experiments 😊

[@Uroojhayat-bc5kj]

I am a MS student and want to do research in Molecular biology and Crisper cas9 is most publishing topic these days thank you Ami li

[@TheRockingBoys207]

ABSOLUTLY MINDDD BLOWNNNNNNN!!!!!!!! i have been learning about CRISPR and im dying to try this experiment!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

[@significantmatters8740]

love how you document your experiment.

[@gregorysmith1134]

It is best to order your plasmid with the promoter, multiple cloning site, antibiotic resistance gene and selectable marker and gene of interest already in it. All you will have to do is make the Agar and carry out the cell transformation.

[@marco-pla]

Very nice experiment, methodological thinking. Well done.

[@r25-j9e]

a new biology professional emerges in the youtube world, waiting for her to spark

[@jessehorstman]

Thank you. Please do it again.

[@DCSTVOFFICIAL]

This is so cool, by the way love your glasses.

[@alexeyeliseykin517]

very cool))) you can try to incubate the transformed cells in liquid LB medium for 1 hour 37°. Then plate these cells on LB agar with antibiotic. (video instructions for transforming the dh5 Alpha are on the NEB website) and you will succeed!!!

[@ld508]

Great video looking forward to a successful update.

[@amymli]

Thank you! I'll give an update here: after a few more days the bacteria did grow, my hypothesis for why it didn't grow as fast as it should have was because I didn't put enough post-CRISPR solution onto the antibiotic plates (aka not following the instructions whoops). If you're planning on trying this out make sure to pour ALL of the CRISPR solution onto the antibiotic plates

[@Jindy2]

This was great! Thanks for sharing your tips on what not to do.

[@amymli]

Thanks for your feedback!

[@jakewarren7826]

It didnt occur to me how accessible this stuff was until I saw someone puriify cas 9 in the lab. I was like "ow yh its just a protein" , very scary but exciting moment :)

[@kaika.hana1]

I just do hope she knows how to properly dispose them that actually includes autoclaving them including all the materials contacted by the bacteria... But yes, CRISPR is a very amazing research you can ever do and know

[@khatharrmalkavian3306]

The thing that freaks me out is that this is not the first KZbin video I've seen of people just ordering gene editing stuff online and splicing things at home. This is probably not a good trend to hand to the internet, which is full of upstanding and well-meaning individuals who would never think of creating a monkey with four @sses.

[@lyon981]

I'm a german student trying to find a good source for my CRISPR/Cas9 Essay. I'm glad I found this video indeed very cool

[@lyon981]

I was originally looking for problems scientists face when trying to transport Cas9 and sgRNA into live organism.

[@mingzegao4794]

Fascinating video!

[@siphokuhleprince4788]

this was amazing🥳🥳🥳🥳next time plz also show us the bacteria under the microscope

[@LM-ch8rh]

Thanks for the video. I'm planning to buy a DIY Crispr kit. Wondering if you were able to get your follow-up experiment to work?

[@amymli]

After waiting for a while, the bacteria did grow. I think the main problem was that I didn't follow the instructions -- I didn't put enough of the transformed bacteria onto the anti-biotic agar 😅 Hope your experient is successful and you learn a lot from it!

[@iraqi3612]

Thank you very much. I wish you good luck in the next experiment. Never forget that a mistake is more valuable than success from the first experience, the mistake teaches you. but the success makes you follow the instructions only.

[@amymli]

100%, thank you so much!

[@somendrapandey1273]

We have to put that mixture in freezer or refrigerator

[@demonye4905]

The only thing i recoginze is E-coil.. the rest is much beyond... It seems at final neither of that the two plates has E.s growing?

[@DavidL-wd5pu]

Great video, science rules!!

[@restrictedendonuclease4340]

See I'm doing my mbbs but I have huge interest in molecular genetics and biotech so what I'm doing is I'm silencing the mRNA molecule in mice which is responsible for formation of myostatin protien which is a negative growth regulator which ..but as lack of apparatus and many things I can't go genome sequencing and other this so please can you tell me another alternative??

[@restrictedendonuclease4340]

And also please tell me habitat of ecoli and ecori plasmid extraction ... Is this have the plandrome GAATTC??

[@farihagul2254]

Could you please tell me where can we get it done? I mean to change eye color or hair color

[@amymli]

To my knowledge I don't think CRISPR can be used for cosmetic/ non-life saving purposes yet, but maybe in the future...

[@Ferdinand7]

It could, but the science (and ethical remorses) are not ready yet

[@theshifzz-u6r]

Your video is just awesomeeeee...!❤✨ But will you please add subtitles in it.

[@QINNIYQ]

Q: So this experiment was to create a strain of ecoli that was resistant to the antibiotic?

[@amymli]

That's correct!

[@sandeepgolui8129]

Excellent try .its inspiring.....great try.. best quality editing...nice to observe this kind of vedio ...thanks a lot madam🇮🇳👍👍🙏

[@MariaFernandazz]

I’m a high school student in Brazil so I don’t really understand synthetic biology very well, would you have a book or online course or whatever other source to help me study more about this subject? I love all of this but I’m having a hard time knowing how to study it by myself. I learn pretty quickly so it doesn’t need to be a very introductory book. It would be extremely helpful. Thanks!

[@amymli]

I've heard that Synthetic Biology: A Primer is a good book to start with. Good luck on your synbio endeavors!

[@randomitems000]

@@amymli 🤙🏾

[@MuhammadAli-oy6zo]

Would love to do at home by myself. Good luck for the next time

[@LAGARTOKID]

Then the bacteria escapes, evolves to a pathological bacteria and boom, you have a bacteria that can't be killed by antibiotics LOL best part is that I still love this video

[@peterxyz3541]

I was pondering the agar from the Chinese grocery store 😂😂😂 instead of pharma grade agar

[@learnbioengieering-general1390]

Can I use crispr cas9 to modify baceria to produce pure meth? Just asking. For science.

[@6root91]

My microbiology senses were tingling throughout the entire video. Still, for a home experiment, it was pretty cool. Makes me wanna purchase some agar plates and a micro-pipette to go with my microscope.

[@novathorn9407]

what is it that was meant to happen and what was it that should have made it happen?

[@leofromthe9159]

I’ve seen a lot of people use crispr on bacteria but I’m curious if it’s possible to edit the genes of a multicellular organisms, such as plants, at home?

[@amymli]

yes! www.the-odin.com/plant-genetic-engineering-kit/

[@AtharvaSalkar-nb7iq]

Are the tips reuseable after boil sterilization

[@andersonchen7501]

Hey Amy! thank you for this great video, I'm a highschooler who wants to conduct similar experiments at home, may I inquire you the total price of those equipments and where you got them from? thank you

[@amymli]

hey! sorry for not seeing this earlier. in total it was ~$200 but you can definitely get more affordable ones at www.the-odin.com/ best of luck!

[@treefrog123a]

Did u kill all nacteria when done

[@ANismo82]

Can you use this to make antibiotics at home?

[@AdrianDucao]

Is the kit from Odin sufficient enough for me to perform a crispr experiment on a mice to make the mice muscular? And what are the repurcusions if i tried it on a human host?

[@amymli]

the Odin only provides kits for bacteria, yeast, fungi, and human cells i believe, if you want to perform a very specific CRISPR experiment, you'd have to design your own g blocks (using software like Benchling) and buy them synthesized from places like Twist or IDT. However, unless you're affiliated with a lab or have professional guidance, I don't recommend trying to go all mad-scientist-y on mammals because although there's a high chance nothing will happen, there's also a chance for off target effects and other things that can go wrong and then it won't be the most ethical scenario you're in

[@gabrieldelmiro8920]

Is it yet possible to look for a CRISPR treatment for a geneticly related disease?

[@amymli]

There are many promising clinical trials currently taking place. I believe just recently the FDA approved of using CRISPR to cure sickle cell anemia (which is very exciting :))

[@gabrieldelmiro8920]

@@amymli and what about diabetes, type 1 or even type 2?

[@suryaprathap2727]

Iam from India can you help to on gene editing on shrimp 🦐🦐

[@simhotlowboatgaming8745]

Can you change a birds color with this kit?

[@fantastico4795]

Y no se contaminó nada 😢??

[@natiedry5470]

Were can I purchase kit like that?

[@amymli]

Right here: www.the-odin.com/diy-crispr-kit/

[@avoniadevile3035]

Thanks for your video l am second year biomedical science student and also got my interest in biosynthetic biology and especially l got an interest in Crispr. Thanks for being honest with mistakes! New subscribe from me, hihi!

[@danbreeden1801]

Thank you for this program

[@Relatista38]

How cool is that? CRIPR at home, hope you make more videos like this :D I am also building some sort of biolab at home xD it will be good if you share some ideas :D TYVM for sharing

[@isaacmadron3916]

I've been want to get into this.

[@NYCdude321]

OMG it's that SIMPLE!??!??!! I'm sure it will succeed if she got a temperature-stabilized incubator.

[@this-abledtheextravertedhe5299]

Fantastic 🤓

[@InquilineKea]

How did you selfie yourself?

[@amymli]

tripod + camera!

[@15youngblack]

What would you recommend studying to get into Crispr?

[@amymli]

just read the paper and google any words/ concepts you don't understand! osu-wams-blogs-uploads.s3.amazonaws.com/blogs.dir/2501/files/2016/11/The-new-frontier-of-genome-engineering-with-CRISPR-Cas9.pdf

[@destroyyourdebt1]

Did you ever figure out the issue?

[@amymli]

The instructions said to put the entire microcentrifuge tube of modified e. coli onto the agar + antibiotic but I only put a few microlitres because I misread. I think that was the problem 😅

[@CanCan-ky9le]

Where can I get Cas9 protein?

[@amymli]

The odin has perishables replacement here: www.the-odin.com/bacterial-crispr-kit-replacement-perishables/ but if you're just looking for the cas9 plasmid you may be able to order it from websites like synthego, thermofisher, twist, amazon, ali express

[@JenembaSquad]

She could have used the spiderman 1 school outing explaination....

[@flareonspotify]

Try making ips cells (turn skin cells into stem cells)

[@__________6597]

When is possible build nanotecnology medicine for kill cancer or infections? Tx

[@amymli]

It is currently possible :)

[@__________6597]

@@amymli in 2021 or 2022?

[@amymli]

@@__________6597 www.cancer.gov/nano/cancer-nanotechnology/current-treatments, I think the problem with cancer is that it's so personalized so even with nanotech delivery, it still may not be effective. If you're interested, I think bioelectricity for curing cancer is a promising approach, check out Michael Levin's work

[@amymli]

But I'm definitely not a cancer specialist at all so take everything I said with a grain of salt

[@__________6597]

@@amymli tx

[@eha521]

what have you learned to do this?

[@sigi_iterator]

YES ANTIBIOTIC RESISTANSE PLAGUE!!!!

[@arabrar1503]

Can't believe she has only 300 subs

[@Matstoen]

Wait a minute, so you finally got this kit that allows you to modify any living organism, and you choose to modify E. Coli? Why?

[@amymli]

They provided e. coli in the kit. It's extremely difficult to get a living organism unless you're associated with a lab

[@nityanandsarkar8054]

WOooow now, that's interesting!

[@mikhalwilliams1933]

Can you make your own crispr kit?

[@amymli]

You can definitely find cheap materials on ali express or other vendors, but I've found that the odin kit has been very thorough in providing & explaining everything you need -- it's also not that pricey! www.the-odin.com/diy-crispr-kit/ they provide extra pipette tips, agar, microcentrifuge tubes, etc. so if you want to perform an additional experiment (ex. express GFP in e. coli) you'd just need to purchase additional biologic material, not everything

[@proxtu1237]

PUEDES SUBTITULAR TUS VIDEOS AL ESPAÑOL POR FAVOR ?

[@InquilineKea]

Macrophage tube????

[@amymli]

a-what now?

[@DevonPhoenix]

Pretending at home just like they do at the lab

[@AcrossTheVoid-ih1jm]

Thank you

[@kothejunglist]

awesome!!!!

[@adespade119]

what could possibly go wrong

[@pedrosalinas4125]

So, you made an antibióticos resistant bacteria? Are you Crazy ?

[@AM-sv2vv]

yaaaas queen

[@aljenembtry7781]

I'm building my own genetics lab at home also!!!!¡¡!!!!!

[@TheOrderoftheSparrow]

Not seeing much of a sterile environment maintenance there. You relying on the antibiotic resistance of your transformants only? Meh guess it’s fine right?

[@tx-sweet-pjg3547]

So where will you turn your mutant hybrids loose ? An ex boyfriend perhaps 🤔😂

[@abhishekadile1270]

So you basically made a anti biotic resistant Bacteria

[@amymli]

Yep, that was the goal!

[@dorfinhorg9458]

Most bacteria we work with in Biotech has some sort of antibiotic resistance. This allows up to grow up the cells we want in media containing antibiotics, which kill off all other contaminants. It is called "selection"

[@israeladedeji4342]

That's good

[@brothermichael2833]

No. That's not what is claimed. I want to see what is being purported. This is insufficient.

[@azukarzuchastux8066]

John 3:16 KJV For God so loved the world, that he gave his only begotten Son, that whosoever believeth in him should not perish, but have everlasting life.