CRISPR CAS9 Gene Editing Technique, The Full principle (Part 2)
ฝัง
- เผยแพร่เมื่อ 28 ก.พ. 2017
- This video is a full explanation of CRISPR-CAS 9 technique and its utilization in gene editing
See also, the principle of CRISPR system:
• CRISPR System and CRIS... - วิทยาศาสตร์และเทคโนโลยี
![[UNCUT] ที่นี่ที่แรก ปู - หาญส์ พูดถึงมหากาพย์หนี้สิ้น I คนดังนั่งเคลียร์ I 10 ก.ค.67](http://i.ytimg.com/vi/KDNA9DZ0p4U/mqdefault.jpg)
Thanks for your clear explanation. However, I think there is one small error. At about 10:25 you say that the gene should be not more than 20bp long. I think what you meant to say is that the gene recognition part of the crRNA is 20bp long. The target gene can be much longer (many 100s or 1000s bp), but the crRNA recognizes a specific 20bp on the target RNA and this is where the CAS9 protein then cuts the target DNA.
This is the best video I have seen on crisper thank you
Thank you so much. I readed a bunch of works trying to understand CRISPR/Cas9 and this helped me to see all clearly. And you even aswered questions I had, like about the PAM sequence. Thank you again
Best explanation of CRISPR Cas9 system in both part.....Thank u.. so much
This is the best explanation about CRISPR I've seen....Thanks a lot
Oh Thank you for your comment, dont forget to subscribe the channel to see all the new videos :)
Hi , thanks for your perfect presentation .. um asking ur permission for using parts of ur presentation for educationsl purposes ..
alaa muayad I agree
Wow! thanks for your brief but huge explanation, keep inspiring!:)
Excellent and lucid presentation of a very complex topic! Kudos!
ITS REALLY BEST VIDEO FOR CRISPR CAS9 EXPLANATION..... REALLY I LIKE IT SO MUCH ..... THANKS A LOT MAM
Thank you alot for explaining in such a simple way 🙏🏼
Love the explaination both part 1 and 2
you are brilliant thank you to the moon and back !!!!! you saved my life
exellent lecture, i think now m clear about CRISPR/Cas9....ver well done
Brilliant explanation I like how smooth the explanation goes
Thank you so much for all your amazing videos! I really appreciate your work :)
Imagine finding a girl like you who is also interested in biology
Best explanation about CRISPR.
Really precise explanation, Thank you very much
Thanks it was very useful and clear , you are an excellent teacher
Wow, how smart is this bacterial system. Thank you for this video ,☑️ goes to show what we are up agenest when fighting these bactras
Great presentation, really helpfull
Excellent explanation! It is great......
thank you for your amazing video. It helps me a lot.
Thanks for this good explanation ;)
thank you for these vidios
Best explanation thanks a lot to make crisper cas9 Best understanding easy
so much helpful. thank you very much.
hi, please explain how do you introduce the PAM sequence into the mouse embryo cells (unless it was introduce artificial prior somehow)? i assume it is a sequence found in the bacterial phage as it is developed as a defence against bacterial virus?
I dont understand why an exonuclease would act on the 3' ends to eliminate the rests of the gene, and then the polimerase will act
Excellent explaination
Hey, Thanks for the explanation, Would you please tell me how the PAM sequence is constructed in mice genome, as you said in the previous video that it only has in the viral genome and bacterial CAS complex can recognise that, so How come it has come in mice genome? Artificially constructed?
very good explanation
thank you for the explanation! one question: 14:31 how could the rest of gene1 be degraded? and how it will exactly end by gene1? thank you
Hi thank you very much for posting and explaining it so well. I have one question. Which paper was the one where they used the blastocyst of the mouse? I need to verryfy the sources correctly for a school project. Thank you very much for your answer :)
it is a great video to understand the technique easily for a beginner like me. thank you a lot and Could you please make other videos for gene insertion and point mutations using CRISPR/CAS9.
Great and simplified explanation of CRISPR CAS technique. Could you make a video for possible unwanted mutations caused by CRISPR and how to prevent it? Is it possible to deliver these plasmids to living patients mutated cells with the same viruses?
Nice explanation. Pls explain gene editing in plants
Awesome!
Thank you very much lover it
Thanks for the good video.
A question: Protospacer Adjacent Motif : What process or processes would I invoke to find the PAM of a totally strange species? What is the name for said process? I don't need to be told how, I want the name of the process. Thanks
Thanks alot.
This was really helpful for my class! I really appreciate this second part just as much as the first.
This is a beautiful explanation! Thank you so much for the two videos, they are great.
Hi, this video is really good. I have one question which will clear my doubt. How does DNA gets chopped out both in case of blunt end or sticky end? Please answer me.
So does double stranded break necessarily mean it's homologous?
You observed that the CRISPR knock-out of protein X either promotes or represses RNA polymerase II transcription, depending on the gene. Which experiments would you carry out to reveal the mechanism for this differential activity?
Hi. Could you please inform me how to find the target DNA in the cell which is knocked out by Cas9 for treating Cancer cells?
SUPER!!
Thank you.
How can I use this technique in viral infection in mammalian cells especially when virus is RNA something like Newcastle disease virus? ???
Nice explanation of CRISPR gene editing. I have one question regarding remaining yellow portion of sequence in lower strand. Is it remaining part of gene one? Could you please explain it?
Thanks a lot
Where is the PAM sequence coming from in the genome of a mouse? In video 1 i think you said PAM is only found in viral genome. Please explain
thanks
?? The rest of gene 1 is removed, the other rest of gene is degraded ?
Who does it and how does that happen..
This is fantastic, thank you! I still have questions though. What exactly is the function of the tracrRNA? I think I heard somewhere that it's to bind Cas9. Also, how is the plasmid processed once it's in the cell? We kinda just saw it in its intact circular form, then suddenly it was broken up.
you should see the first part of the video
The cell's machinery replicates (depending on the strength of the origin of replication) and transcribes the plasmid once it's on the inside. You should watch and read other sources about this topic.
Why the type 2 system is mostly used??? Plzzzz answer me
thank you so much for explaining it so well. Among all the videos I have seen, I feel that your one is the best. I have two questions. 1) do we know previously which position the CAS9 is going to do dsbreak? 2)how do you design template strands when you are planning to KO a certain gene? does that mean the template strand has some mutations? as result, it's not going to translate the protein. Thank you so much!
1 PAM
2 Yes, CRISPR-CAS9 is aimed at preventing genetic diseases at embryonic level. The template to be KO'd has a mutation and so a 'healthy' version of the template is also provided as a replacement.
Hope this helps.
14:07 how does rest of the gene get degraded? Where and how does it stop?
At 10:20 you talked about how the Enzyme can be modified to recognise the PAM. How does that happen? Also, can the converse of this process take place? As in, instead of knocking out an undesired gene, we add a desired one?
Perhaps by replacing the template with the saud gene or some other technique?
For the first question: what she meant was that there are multiple bacterial organisms that have different pam recognizing site that you can use based on your gene of interest.
2nd: it is possible to add your desired gene by having the dna template to be a translatable gene so that your protein of interest can be expressed.
How rest to gene is being chopped off, pls explain?
How is PAM sequence present in eukaryotic system?
I do not understand how after double strand break caused by Cas9 enzyme, the rest of the gene will be degraded before he new DNA gene in ligated and who grantees the complete degradation (why exactly and only the gene is degraded, why does not the degradation go beyond the gene or only take a part of the gene
What about the rest of Gene 1 when finished with the sticky end synthesis? it was never fully knocked out
Yes, I have the same confusion.....
It would be interesting to carry this further where you discuss things like site directed mutations as a precision tool for gene editing, for example. Really outstanding! Who are you? ;)
HAHA, thank you for your comment. I will try to make a video about your suggestion. I am NOONE loool :D
hahaha that makes two of us.
HDR does not use dna template while nhej uses template???
🔥🔥🔥
I think NHEJ and HDR are mixed up in the video...which is actually a very big mistake.... This makes me question the validity of this channel, which is sad because I really liked the animation and stuff.... Also, where does gRNA come into play? I'm just starting out on CRISPR
I agree ! She completely mixed up NHEJ and HDR !!!
Wrong explanation for DNA repair machinery both for NHEJ and HDR
There are a lot of things explained well in this video so the validity of the channel does not need to be questioned. This kind of attitude will keep people from adding value to the scientific community. One mistake does not put every single other piece of work into question. Overall it is a great video and I guess it has helped many people. Giving constructive feedback and helping people to improve for the future is one thing. But this sort of criticism will stop other people from adding value and will not help anyone in the long term.
@@stefanieginster6099 Wow science is a process which totally depends on challenging and questioning. It's our quality control. When does anyone ever say in science "This does not have to be questioned"? As if challenging someone is an "attitude"! WTH
You should definitely clarify 'DNA repair template' thing.
Is there an age limit for crispr? I'm a 39 year young soldier stationed in Hawaii and I just discovered that I have marchado Joseph disease. This news has destroyed my marriage, my employment. I will be med boarded because of this. Please help.
Why, and how, is Gene 1 removed in the process?
The way of your presantation is very well but, I have some doubts on DNA repair machanism which you deal..
DNA repair is a huge topic, I cannot mention it all in the video, maybe I can do another video speaking specifically about DNA repair :)
Not clear and not correct...
i love you! i have a proyect in way of this, actually i gotta take it to revision this tuesday , thank you so much!! note: could you please give a message that contains the information references for this video??
Hey, thank you for your comment. Actually, I read many papers to come up with this video and it was a couple of months ago so it is hard now to collect all of them, but this is one of them, you can read it, it contains good information as well:
www.neb.com/tools-and-resources/feature-articles/crispr-cas9-and-targeted-genome-editing-a-new-era-in-molecular-biology
thank you so much :D
I know this was 3 years ago, but for other people interested: www.ncbi.nlm.nih.gov/pmc/articles/PMC4385387/
This reference was very useful (accompanied by this video, of course).
the way she said, ''we don't want this gene'', quite interesting
Complete and detailed explanation of crispr gene editing but the only thing you need to improve is fluency while talking although loved your content
Thank's for this very good video. How we can use this technic in lab? Thats possible to do in common labs or want special equipment?
You can perform it in the lab in an Eppendorf tube. You need your cells, the crRNA, the tracrRNA, the CAS9 enzyme, and the DNA template (watch min 4.09). You can buy all of these.
Thank you. Do you work on crispr technic in lab?
Very good video mam but DNA repair is not very clear. In homologus you have denatured all yellow part( Target gene) And in second you hv just denatured one part.
the yellow part remain as it is.
@12:18 representation of sticky ends is wrong.
at last slide
Shaista video
DNA should never be written 3' to 5', always 5' to 3'.
How did the CRISPR-Cas get into the eukaryotic cell?
plasmid
but there is no PAM in eucariotic cells ??
wasnt it only in virus dna?
why is PAM
in mouse dna ?
you make it more confusing because of too many pauses, rest is good! great work
Chimera Virus