at 6:27, I think we can also clone the Sp Cas9 plasmid with Neomycin / Puromycin resistance sequences. That would essentially mean we wouldn't have to rely on FACS and PCR further down the steps, and can essentially select the transformants right from the cell culture dish while comparing it with a control of non-transformants. Please let me know if I'm correct.
@@animatedbiologywitharpan thanks for the clarification. Great video as always! How the procedure is performed in the lab is exactly what i was looking for, as I'm about to start working with crispr to understand the role of a transcription factor soon
Nice demonstration. My question is why not label every plasmid (sgRNA, Cas9)) with different fluorescent protein (green yellow, red, etc) and sort the cells with two fluorescent proteins and expand from there ? This saves time and effort ! Also you can use antibiotic resistance.
thanks for the question in reality there might be 300 clones (not 3) which you have to screen for identifying your mutant.....so it becomes hectic in my opinion.
@@animatedbiologywitharpan You have provided one clear video. thanks. I have a question about your answer. When you say "3 clones" are you referring to the three plasmids in the video? If that is the case, I would imagine multiple 96 well plates would be needed to yield one cell line. How long would the entire process take (idea to success?) for a two grad student team?
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I have few question How far we are from these technologies, i mean precision medicine? Who will able to afford it rich or poor? The way climate changes occuring so fast will it be able to inbuilt these precision medicine in particular individual? To make precision medicine how much time is needed for one person, average time?
Other answers please Can u answer them Who can afford it? How much average time it will take to make precision medicine for individual person?@@animatedbiologywitharpan
Good question .....photoreceptors are differentiated cell so doing crispr is bit difficult...but doing crispr editing in stem cell then differentiating them into photoreceptor is easily doable
@@animatedbiologywitharpan That makes sense! I have some more questions if you could help me 😅 So in university we were asked to 'build' a gene therapy and we thought about doing gene editing with Crispr ex vivo. We would collect capillary cells and reprogram them into stem cells, then do the gene editing with Crispr, then differentiate them into photoreceptor cones and finally transplant these cells into the patient eye (the patient had photoreceptor cones degeneration because of a CNGB3 mutation). Is this transplatation even possible?
I know this is an old comment but there is currently cutting edge research delving into exactly this. Yes, auto-immune diseases are due to a genetic factor in immune cells, which causes them to fight the body. CRISPR and gene therapy have already been used in humans with autoimmune disease with success! Though it is still in the research stage. This is a very rough explanation as I am not a gene scientist.
Jesus is the son of God who died and rose three days later so you can be free forgiven healed delivered saved baptized and so much more. Believe in Him as the son of God and you are saved eternally and will live eternally. He's awesome and wants us knowing Him and spending time with Him. Ask anything in Jesus name and He will do it.
@@haileybaker8542 no offense but I can post a comment about anything I want to say where I want to say it. Freedom of speech allows me and you and everyone that same right. Also Christians are supposed to evangelize the bible everywhere and this was part of my everywhere the day I wrote it. Everyone needs Yeshua and needs to know about God..there's more to God than my comment. God bless you 🙏💖
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Great video, thanks for sharing. One of the few I've seen about the actual workflow of CRISPR edits.
Could you recommend any other RUclips videos that have proper explanation of the protocol?
A very informative Video. Thank you for clearly explaining, in a easily understandable manner for a beginner like me.
Please share among your friends and help me to reach big audience
Animated biology With arpan how can I get crisp case 9
at 6:27, I think we can also clone the Sp Cas9 plasmid with Neomycin / Puromycin resistance sequences. That would essentially mean we wouldn't have to rely on FACS and PCR further down the steps, and can essentially select the transformants right from the cell culture dish while comparing it with a control of non-transformants. Please let me know if I'm correct.
I completely agree
@@animatedbiologywitharpan thanks for the clarification. Great video as always! How the procedure is performed in the lab is exactly what i was looking for, as I'm about to start working with crispr to understand the role of a transcription factor soon
Nice demonstration. My question is why not label every plasmid (sgRNA, Cas9)) with different fluorescent protein (green yellow, red, etc) and sort the cells with two fluorescent proteins and expand from there ? This saves time and effort ! Also you can use antibiotic resistance.
Pretty nice comment. which steps exactly would you be doing this ? and is it more economical than this method ?
Спасибо!
Very nicely presented. Thanks.
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@@animatedbiologywitharpan Most definitely. Your channel content is really interesting and informative. ✌️✌️👏👏
at around 9:48 you said it is tedious. How long does it take to run PCR on all three "target colonies". What part is the most repetitious and tedious?
thanks for the question in reality there might be 300 clones (not 3) which you have to screen for identifying your mutant.....so it becomes hectic in my opinion.
@@animatedbiologywitharpan You have provided one clear video. thanks. I have a question about your answer. When you say "3 clones" are you referring to the three plasmids in the video? If that is the case, I would imagine multiple 96 well plates would be needed to yield one cell line. How long would the entire process take (idea to success?) for a two grad student team?
Great informative video!! made it very simple and easy to digest.. cheers!
Please share my channel link with your friends and help me to reach big audience
Great information
In order to make high quality content consistently, we need support from you. Please support us by using super thanks option. Super thanks icon is present below the video ( a heart sign with $ in it ) . You can support using paytm/ phone pe/ gPay / paypal. Your small contribution means a lot for us.
Many thanks for your video. Could you please share with us which tools did you use to make this animation?
I use PowerPoint only
@@animatedbiologywitharpan Thanks again, for your reply!
he has used NCH software
thank you very much
You are welcome
I have few question
How far we are from these technologies, i mean precision medicine?
Who will able to afford it rich or poor?
The way climate changes occuring so fast will it be able to inbuilt these precision medicine in particular individual?
To make precision medicine how much time is needed for one person, average time?
Quite far from precision medicine
Other answers please
Can u answer them
Who can afford it?
How much average time it will take to make precision medicine for individual person?@@animatedbiologywitharpan
what are the three ways to upregulate the expression of a gene in arabidopsis using the CRISPR/Case9 method...pease sir help me in this
Read this article to learn more www.nature.com/articles/s41587-019-0296-7
Hey Arpan! I really like your graphics! Where do you find them? Especially the ones in slides 4-6?
I have done it in Adobe illustrator then used photoshop....but believe me same graphics can be generated using Microsoft power point
I provide tution for this as well over zoom
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So what are the long term effects....will it reverse, mutated or evolve in the future?
CRISPR induced mutations are stable
@@animatedbiologywitharpan Thank you for educating me and replying so quickly!
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@@animatedbiologywitharpan Already have done soo!!
@@animatedbiologywitharpan ..... how do you know ... ??... monkeys playing with bombs....
Can you share a paper or source for transcriptional activation repression of RNA by crispr cas9.
Hi!! Could this technique be possible to use in photoreceptor cones?
Good question .....photoreceptors are differentiated cell so doing crispr is bit difficult...but doing crispr editing in stem cell then differentiating them into photoreceptor is easily doable
@@animatedbiologywitharpan That makes sense! I have some more questions if you could help me 😅
So in university we were asked to 'build' a gene therapy and we thought about doing gene editing with Crispr ex vivo. We would collect capillary cells and reprogram them into stem cells, then do the gene editing with Crispr, then differentiate them into photoreceptor cones and finally transplant these cells into the patient eye (the patient had photoreceptor cones degeneration because of a CNGB3 mutation). Is this transplatation even possible?
@@catarinabarbosa9471 my email is arpanparichha1994@gmail.com ...feel free to connect
@@animatedbiologywitharpan Done so! Thanks!!
I have replied all ready, if you like the content please share with your friends and help me to reach big audience
At what stage of cell can this Technology be used?
Cells that are growing
Gracias!
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Dada I'm from, kolkata and I am in class 10th, I'm looking forward to start a startup based on Crispr... So I wanted to ask you. Is this idea good??
Yes why not...it's going to bloom in future
what chemicals are used?
Can it cure autoimmune diseases ?
I know this is an old comment but there is currently cutting edge research delving into exactly this. Yes, auto-immune diseases are due to a genetic factor in immune cells, which causes them to fight the body. CRISPR and gene therapy have already been used in humans with autoimmune disease with success! Though it is still in the research stage. This is a very rough explanation as I am not a gene scientist.
which microscopy are you using?
Zeiss discovery sterozoom fluorescence microscope is very good for the screening purpose
Great channel
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@@animatedbiologywitharpan Crispr-cas9 can cut faulty gene in genetic genes disease like polycystic kidney disease sacs of fluid. Thanks
Good job
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When we expect this treatment for muscular dystrophy treatment
It's a long path as of now
@@animatedbiologywitharpan any treatment for limb girdle muscular dystrophy treatment sir in the world with any cost
make my hair grow again??
Following
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I can't understand his accent. ;)
Jesus is the prophet of God not son of god
Jesus is the son of God who died and rose three days later so you can be free forgiven healed delivered saved baptized and so much more. Believe in Him as the son of God and you are saved eternally and will live eternally. He's awesome and wants us knowing Him and spending time with Him. Ask anything in Jesus name and He will do it.
What does that have to do with the video?
@@haileybaker8542 no offense but I can post a comment about anything I want to say where I want to say it. Freedom of speech allows me and you and everyone that same right.
Also Christians are supposed to evangelize the bible everywhere and this was part of my everywhere the day I wrote it. Everyone needs Yeshua and needs to know about God..there's more to God than my comment. God bless you 🙏💖
@@SherriPJesus L
can i become spider man by genetically modify myself??????
No