Hey everyone! If you're interested to learn more about CRISPR, we'd love for you to check out our new 4-week CRISPR Crash Course. It's completely free to join and we're excited to hear what you guys think of it! Sign up at: info.abmgood.com/crispr-crash-course
you prolly dont care at all but does anyone know a trick to get back into an Instagram account..? I somehow lost my login password. I would love any assistance you can give me.
@Brodie Enrique thanks so much for your reply. I got to the site through google and im trying it out now. I see it takes a while so I will get back to you later with my results.
Start off your new year with a brief introduction to the most revolutionary genetic engineering tool available today - CRISPR/Cas9 - th-cam.com/video/1aJxXWkE3Ek/w-d-xo.html
Thanks for your comment, Ragnar -- glad you enjoyed the video! Don't hesitate to post any questions you have, here. Or, if you would like more information, you can check out our knowledge base: goo.gl/df55ap
I'm having some issues with the basic CRISPR mechanism in bacteria: 1. "The invaded cell recognises the foreign DNA and stashes away a snippet of it in the CRISPR structure". So if the cell can do that, why can't it simply chop it up and be done with it? 2. How does the cell survive the initial attack, whereby gazillions of virus copies are created and then expelled from the cell, usually accompanied by cell death due to membrane rupture? 3. How does the RNA template inside Cas9 mate up with the DNA? RNA has uracil instead of thymine! And for genome editing, Cas9 requires a PAM match before it attempts an RNA match. This would appear to be a very strong restriction on where edits can be made. So 4. Can the Cas9 PAM be made programmable, thus allowing edits to be made anywhere?
+Andrew Palfreyman I will try to answer your questions one by one here: 1) The idea of the CRISPR System is to act as an immune system - in our bodies after an infection the body stores a copy of the antibody secreting B-Cell (memory B-Cells) so that in case of an infection with the same agent in the future it can react quicker in clearing it (therefore, you won't experience any symptoms from the same agent again). the CRISPR System acts in a very similar way. 2) typically this type of system doesn't work on the first bacteria that gets infected. After the initial infection and burst some of the un-packaged viral DNA also gets released into the extra-cellular environment. It is this DNA that the other bacteria can take up and incorporate into their CRISPR System and protect themselves from the virus. 3) the same way we have transcription. In transcription a copy of the DNA is made into RNA. U just binds to A. 4) the restriction isn't as big as you think, the PAM sequence is NGG which is a very common element found in the genome. However, this PAM sequence is indeed programmable in the sense that different Cas9 proteins recognize different PAM sequences - you can check out our knowledge base for all the details: goo.gl/ceUmEE please let us know if you have any further questions!
Thank you. I'm doing a project on CRISPR-Cas 9 in the role of drug discovery. Can you explain further how personalized medicine can be made from this system?
Sarah Ng This system is the most versatile tool in genome editing - it allows for genetic modification simply by designing a 20bp guiding sequence. Given its simplicity, CRISPR/Cas9 has the potential to modify deleterious disease causing mutations in an individuals DNA. Although this system as been used successful in many model organisms ( goo.gl/ceUmEE ) recent research has raised considerable debate about its effectiveness and ethical use: ow.ly/M10b2
Thanks for very nice and well demonstrated video lecture on CRSIPR Cas-9 mechanism and its novel applications. Once a Bacterium is infected by a Bacteriophage, what type of enzymes cleave the viral genome within bacterium, to be remembered by Bacterial genome for recognition in future by CRISPR-Cas9 complex. I mean a detailed mechanism of Adaptive immune system adapted by Bacteria.
Hi Rakeeb, Glad you enjoyed our video! In the bacterial immune system, invading foreign DNA (such as bacteriophage DNA) is cleaved by Cas nucleases. You can read more details about the mechanism here: goo.gl/df55ap
Can I know if, this video is still accurate in terms of CRISPR development untill today. I mean is this all, or it has grow to have more systems than the 3 shown in the video.
Hi Andrew, thanks for commenting! Any new technology must certainly be used with caution and assessed rigorously for any associated environmental risks. CRISPR Cas9 is mostly used in controlled settings for life sciences/cell biology research. There have certainly been some concerns regarding the use of CRISPR in creating transgenic organisms that are mixing with "wild" populations (there's a great nature article here summarizing CRISPR applications and government concerns: goo.gl/vlKWoz). As such, researchers have invented technologies such as "Gene Drives/Reverse Gene Drives" (see discussion about transgenic mosquitoes) that can provide this much needed control.
Hi Kushagr, it sounds like an interesting project - we wish you the best in your research endeavors! If you need any help carrying out your gene editing projects, we do have products and services that might help you fast-track the process, feel free to check it out: goo.gl/9VHtk7
To better illustrate this, I would like to refer you to this image from this Nature paper: www.nature.com/articles/srep40638/figures/4 Position 1 is closest to the PAM sequence (3'end of target sequence), while Position 20 is furthest away from the PAM sequence (5'end of target sequence). Since the 3' end of the target sequence is known as the Seed region, which determines the binding and cleavage efficiency of Cas9, any mismatches here will abolish Cas9 activity at this site. If the mismatches were closer to the 5' end, the cleavage efficiency may be reduced, but there may still be some Cas9 activity at this site (off-target cleavage).
+riu sengupta Thank you for your question! In theory, yes, it should, however, we have not come across any publications citing the use of CRISPR/Cas9 system on quadruplex DNA.
hello, i got a question now, i am an univrsity student, i am doing my finial year project, which is about genetic modifing the HCMV, for the honologous recombination, i have to add the specific template into the target ( the double strand DNA break site), but i would like to ask if i buy the template sequence from science company, i need to amplify the number of template sequence, which is inserted into the plasmid, i need to add primer for the PCR, but i read a paper for the similar experment, they are using both of the forward and reverse primer for the PCR, thus i would like to ask why do i need the verse primer for the PCR???
Hi Asit! Thanks for your comments. In the bacterial immune system, invading foreign DNA is cleaved by Cas nucleases which are then captured and integrated into the CRISPR locus as what we call spacer sequences. If the same foreign DNA enters the cell, these spacer sequences are used as templates to express the RNA that guides Cas nucleases to the foreign DNA for degradation. If you're interested to learn more, we have an entire knowledge base covering the CRISPR Cas9 and how to do gene editing using this system: goo.gl/df55ap
Thank you for your quick reply. Can you please explain the mechanism through which the cas nuclease cleave the foreign DNA first time before it generate the spacer sequence.
Hi, This is a very promising concept. Is it a theoretical/Laboratory work? OR is there a medical procedure available for patients? I will be very thankful. I have ankylosing spondilitis and HLAB27+ Thanks
+Tushar Goel This technology is still in development. Many researchers around the world have taken an interest in this technology since it offer a simple solution to Genetics Engineering. However, we are still far away from being able to use this technology safely in a clinical setting. Many issues such as the off-target mutations caused by this technology still needs to be addressed. There are companies such as Editas Medicine (editasmedicine.com/index.php) who are looking to use this technology in therapeutics.
Mount -ain You could find all our sources at the very bottom of our knowledge base: www.abmgood.com/marketing/knowledge_base/CRISPR_Cas9_Introduction.php
Cas9 Nickase cuts only one strand of DNA, unlike Cas9 Nuclease which cuts both strands of DNA. So, in order for editing to occur, you need to use two sgRNAs targeting two target sites near each other, one for each strand of DNA to be cut. If only one sgRNA is used with Cas9 Nickase, the single stranded cut (nick) will be repaired, and no editing will occur. This reduces the chance of off-target effects because the chances are very low that both of your target sites will occur in close proximity to each other in some other place in the genome. Hope this helps!
Hugo090519 Be sure to subscribe to our channel to be the first to know when we upload our next video! We have some great video's lined up including continuation of our current series (i.e. CRISPR Cas9 and AAV) as well as starting new series on Next Generation Sequencing (NGS) and PCR!
I plan to perform a gene knockout on a specific gene. Since bothe NHEJ and HDR can be used for gene knock-out, then which kind of method should I choose? why?
+Wenfang Xie Thank you for your question! You can certainly use both the NHEJ and HDR pathway for gene knock-out, however, HDR is more commonly used to make precise modifications (i.e. specific nucleotide changes). This is because in order to utilize HDR, you will also have to include a "repair template" containing the desired sequence as well as additional homologous sequence immediately upstream and downstream of the target. You will also have to keep in mind that the efficiency of HDR is generally low (
Hi Adila, thanks for leaving your comment and reaching out to us! We're glad our graphics are useful to you. Please contact info@abmGood.com with your inquiry and a list of the images you'd like to use and someone from our team will get back to you, shortly!
Will a single gRNA work for all the genes, as in the all in one vector? What are your thoughts on the use of two targets for the same gene to delete the intervening sequence?
Rachel Jesudasan Different gRNA's have different targeting efficiency, and so it is hard to predict how well a given gRNA will perform. Therefore, it is always ideal to design 2-3 different gRNA constructs against your desired gene and use them individually as well as grouped together to increase the efficiency of gene knockout. The all-in-one system provide an ideal tool for transfecting both the gRNA and the Cas9 protein with a single transfection vector which increases the transfection efficiency. This tool is especially useful when you already had success with a particular sgRNA sequence. If you will be using two or more gRNA to delete large portions of the genome it is critical to ensure that off target effects of the gRNA are minimized as it could potentially lead to unwanted off-target effects. for more information check out our latest video at: th-cam.com/video/dXPDefej0Ps/w-d-xo.html
Very soon we will be releasing our next video in this series which will walk you through the gRNA design procedures. Briefly here, we would recommend you to design your gRNA against exon 1 or 2 of the dystrophin gene; this will ensure that frame shift mutations caused by NHEJ do not lead to partially functional gene product. you could also try designing multiple gRNAs against the dystrophin gene and transfect all of them together. abm also provides these gRNA pre-designed and you could find them here: Human- www.abmgood.com/DMD-CRISPR-Lentivector-K0609305.html; Mouse - www.abmgood.com/DMD-CRISPR-Lentivector-K4939705.html; Rat - www.abmgood.com/DMD-CRISPR-Lentivector-K6920905.html
Yes, it can! CRISPR can be used to knock-in any gene, and opsins are no exception. Here is an example of a paper that knocked in a CAG-LSL-ChR2-Tdtomato cassette: www.sciencedirect.com/science/article/abs/pii/S1534580718303277 The most common combination of CRISPR and optogenetics, however, is to create a photo-activatable Cas9. There's a good blog post about some of these photo-activatable CRISPR tools here: blog.addgene.org/optogenetics-crispr-using-light-to-control-genome-editing.
thelongboardsling yes it will work on extracted DNA as well. The requirements will be a functional Cas9 protein and a full length gRNA (including the target sequence and the scaffold sequence). In fact there has been some recent research on this - in order to use Cas9 to perform Restriction digests and Gibson assembly (goo.gl/PQAqae)
Hi Zia, thanks for watching our video and leaving a comment! I believe the biophysics of the mechanism are still being heavily studied, however, what is somewhat known is that an "R loop is formed" in which the sgRNA intrudes upon the DNA double helix to form an RNA-DNA hybrid helix with the target strand and displacing the opposite nontarget strand (from this paper: goo.gl/3vB4O9).
Hi Trafalgar - this was one of our earlier videos. Our newer CRISPR videos have much better sound quality! You can get our entire playlist here: goo.gl/eaxKXK
Does any University that does these studies and experiments have a volunteer program for humans, say you have a cancer in the brain where there nothing that can be done, I wonder if this would be helpful in a cure while still alive.
+Baby Boo . This technology is very new. We are still a long way from being able to use this technology in a clinical setting. There are researchers that are working towards that goal such as Editas Medicine (editasmedicine.com/index.php).
Thank you for the answer, My pastor has a son that is 15 and he has a Very rare brain tumor[cancer] was thinking this would be great if it could be done as you say in a clinical setting, they have done all they can. Thank you again for the answer.
+Baby Boo -You might want to tell your pastor to check out Dr. Burzynski's antineoplastons protocol, which has helped and healed many with 3rd and 4th stage brain tumors. Hope this helps...
I do know of Dr. Burzynski's and he is one of the best I believe my dad has a book/articles on or about him and we have a friend who has cancer,. I do not have any health problems but I was just wondering if any if these Universities do experiment type studies.
+Sherry Von We are sorry to hear that, if you like us to explain anything in more detail please let us know! We will either answer you here or make another video to go into that detail! also be sure to check out our knowledge base for more detailed information: goo.gl/ceUmEE
Wow. The narrating voice in this video somehow really irritates me, the pitch of it really gets into my bones. To bad for me, because it seems like a really good video.
Hi APOLOnl, thanks for your comment! Sorry you didn't like the narration, our future videos have better sound quality. If it helps, you can always mute the video and play it with our closed-captioning turned on. We also have a comprehensive knowledge base where you can find all this information and more: goo.gl/df55ap.
Hey everyone! If you're interested to learn more about CRISPR, we'd love for you to check out our new 4-week CRISPR Crash Course. It's completely free to join and we're excited to hear what you guys think of it! Sign up at: info.abmgood.com/crispr-crash-course
you prolly dont care at all but does anyone know a trick to get back into an Instagram account..?
I somehow lost my login password. I would love any assistance you can give me.
@Maverick Royce instablaster :)
@Brodie Enrique thanks so much for your reply. I got to the site through google and im trying it out now.
I see it takes a while so I will get back to you later with my results.
@Brodie Enrique It did the trick and I actually got access to my account again. I am so happy!
Thanks so much you really help me out !
@Maverick Royce no problem :)
Start off your new year with a brief introduction to the most revolutionary genetic engineering tool available today - CRISPR/Cas9 - th-cam.com/video/1aJxXWkE3Ek/w-d-xo.html
An excellent video! Saved me at least one hour of reading in 6 mins.
Thanks for your comment, Ragnar -- glad you enjoyed the video! Don't hesitate to post any questions you have, here. Or, if you would like more information, you can check out our knowledge base: goo.gl/df55ap
This is the best video on CRISPR system, thanks a lot!
uhm, i've watched more than 20 and this one doesn't make the top 10. Look again
can you tell me which one was the best?
I'm having some issues with the basic CRISPR mechanism in bacteria:
1. "The invaded cell recognises the foreign DNA and stashes away a snippet of it in the CRISPR structure".
So if the cell can do that, why can't it simply chop it up and be done with it?
2. How does the cell survive the initial attack, whereby gazillions of virus copies are created and then expelled from the cell, usually accompanied by cell death due to membrane rupture?
3. How does the RNA template inside Cas9 mate up with the DNA? RNA has uracil instead of thymine!
And for genome editing, Cas9 requires a PAM match before it attempts an RNA match. This would appear to be a very strong restriction on where edits can be made. So
4. Can the Cas9 PAM be made programmable, thus allowing edits to be made anywhere?
+Andrew Palfreyman I will try to answer your questions one by one here:
1) The idea of the CRISPR System is to act as an immune system - in our bodies after an infection the body stores a copy of the antibody secreting B-Cell (memory B-Cells) so that in case of an infection with the same agent in the future it can react quicker in clearing it (therefore, you won't experience any symptoms from the same agent again). the CRISPR System acts in a very similar way.
2) typically this type of system doesn't work on the first bacteria that gets infected. After the initial infection and burst some of the un-packaged viral DNA also gets released into the extra-cellular environment. It is this DNA that the other bacteria can take up and incorporate into their CRISPR System and protect themselves from the virus.
3) the same way we have transcription. In transcription a copy of the DNA is made into RNA. U just binds to A.
4) the restriction isn't as big as you think, the PAM sequence is NGG which is a very common element found in the genome. However, this PAM sequence is indeed programmable in the sense that different Cas9 proteins recognize different PAM sequences - you can check out our knowledge base for all the details: goo.gl/ceUmEE
please let us know if you have any further questions!
How is it that digestion of first viral genome takes place...
Great! Thank you!
Thank you very much Doctor but there is a bit of a problem with the sound
Thank you. I'm doing a project on CRISPR-Cas 9 in the role of drug discovery. Can you explain further how personalized medicine can be made from this system?
Sarah Ng This system is the most versatile tool in genome editing - it allows for genetic modification simply by designing a 20bp guiding sequence. Given its simplicity, CRISPR/Cas9 has the potential to modify deleterious disease causing mutations in an individuals DNA. Although this system as been used successful in many model organisms ( goo.gl/ceUmEE ) recent research has raised considerable debate about its effectiveness and ethical use: ow.ly/M10b2
Thanks for very nice and well demonstrated video lecture on CRSIPR Cas-9 mechanism and its novel applications.
Once a Bacterium is infected by a Bacteriophage, what type of enzymes cleave the viral genome within bacterium, to be remembered by Bacterial genome for recognition in future by CRISPR-Cas9 complex. I mean a detailed mechanism of Adaptive immune system adapted by Bacteria.
Hi Rakeeb,
Glad you enjoyed our video!
In the bacterial immune system, invading foreign DNA (such as bacteriophage DNA) is cleaved by Cas nucleases. You can read more details about the mechanism here: goo.gl/df55ap
Thanks a lot for reference and supporting material...
Can I know if, this video is still accurate in terms of CRISPR development untill today. I mean is this all, or it has grow to have more systems than the 3 shown in the video.
Great video, thanks!
hey, just wanting to know if there are any environmental risks to this procedure? great informational video though
Hi Andrew, thanks for commenting! Any new technology must certainly be used with caution and assessed rigorously for any associated environmental risks. CRISPR Cas9 is mostly used in controlled settings for life sciences/cell biology research. There have certainly been some concerns regarding the use of CRISPR in creating transgenic organisms that are mixing with "wild" populations (there's a great nature article here summarizing CRISPR applications and government concerns: goo.gl/vlKWoz). As such, researchers have invented technologies such as "Gene Drives/Reverse Gene Drives" (see discussion about transgenic mosquitoes) that can provide this much needed control.
Wondering if it can change the DNA? And if the DNA is replaced by the synthetic RNA?
How long will it be before it's available for the public so I can cure diseases and help people walk again I would appreciate an answer thank you
You guys are awesome!.
Glad you found this video helpful! :)
just started working on CRISPR CAS for generating low phytate soybean
Hi Kushagr, it sounds like an interesting project - we wish you the best in your research endeavors! If you need any help carrying out your gene editing projects, we do have products and services that might help you fast-track the process, feel free to check it out: goo.gl/9VHtk7
Excuse me... what do you mean with "Since mismatches at the 5' end of the gRNA are tolerated"?
To better illustrate this, I would like to refer you to this image from this Nature paper: www.nature.com/articles/srep40638/figures/4
Position 1 is closest to the PAM sequence (3'end of target sequence), while Position 20 is furthest away from the PAM sequence (5'end of target sequence). Since the 3' end of the target sequence is known as the Seed region, which determines the binding and cleavage efficiency of Cas9, any mismatches here will abolish Cas9 activity at this site. If the mismatches were closer to the 5' end, the cleavage efficiency may be reduced, but there may still be some Cas9 activity at this site (off-target cleavage).
This is excellent genetic research. Thanks.
No problem! Thanks for watching! :)
Would a CRISPR/Cas9 mechanism work efficiently in quadruplex DNA or any DNA sequence having a non-canonical or secondary structure?
+riu sengupta Thank you for your question! In theory, yes, it should, however, we have not come across any publications citing the use of CRISPR/Cas9 system on quadruplex DNA.
very well explained video
hello, i got a question now, i am an univrsity student, i am doing my finial year project, which is about genetic modifing the HCMV, for the honologous recombination, i have to add the specific template into the target ( the double strand DNA break site), but i would like to ask if i buy the template sequence from science company, i need to amplify the number of template sequence, which is inserted into the plasmid, i need to add primer for the PCR, but i read a paper for the similar experment, they are using both of the forward and reverse primer for the PCR, thus i would like to ask why do i need the verse primer for the PCR???
Can you please explain, how this system is helpful in bacterial immune system.
Hi Asit! Thanks for your comments. In the bacterial immune system, invading foreign DNA is cleaved by Cas nucleases which are then captured and integrated into the CRISPR locus as what we call spacer sequences. If the same foreign DNA enters the cell, these spacer sequences are used as templates to express the RNA that guides Cas nucleases to the foreign DNA for degradation. If you're interested to learn more, we have an entire knowledge base covering the CRISPR Cas9 and how to do gene editing using this system: goo.gl/df55ap
Thank you for your quick reply. Can you please explain the mechanism through which the cas nuclease cleave the foreign DNA first time before it generate the spacer sequence.
Hi
What’s the best references for crisper cas 9 at the moment
Hi, This is a very promising concept.
Is it a theoretical/Laboratory work? OR is there a medical procedure available for patients? I will be very thankful. I have ankylosing spondilitis and HLAB27+ Thanks
+Tushar Goel This technology is still in development. Many researchers around the world have taken an interest in this technology since it offer a simple solution to Genetics Engineering. However, we are still far away from being able to use this technology safely in a clinical setting. Many issues such as the off-target mutations caused by this technology still needs to be addressed. There are companies such as Editas Medicine (editasmedicine.com/index.php) who are looking to use this technology in therapeutics.
Very well explained, Can you please give your source of information so I can use some of this material and have the chance to reference it. Thanks.
Mount -ain You could find all our sources at the very bottom of our knowledge base: www.abmgood.com/marketing/knowledge_base/CRISPR_Cas9_Introduction.php
could anyone explain me how do the cas9nickases to reduce the off-targets? i didn´t understood ( ..im not english speaker)
Cas9 Nickase cuts only one strand of DNA, unlike Cas9 Nuclease which cuts both strands of DNA. So, in order for editing to occur, you need to use two sgRNAs targeting two target sites near each other, one for each strand of DNA to be cut. If only one sgRNA is used with Cas9 Nickase, the single stranded cut (nick) will be repaired, and no editing will occur. This reduces the chance of off-target effects because the chances are very low that both of your target sites will occur in close proximity to each other in some other place in the genome. Hope this helps!
Thanks for posting
Hugo090519 Be sure to subscribe to our channel to be the first to know when we upload our next video! We have some great video's lined up including continuation of our current series (i.e. CRISPR Cas9 and AAV) as well as starting new series on Next Generation Sequencing (NGS) and PCR!
I plan to perform a gene knockout on a specific gene. Since bothe NHEJ and HDR can be used for gene knock-out, then which kind of method should I choose? why?
+Wenfang Xie Thank you for your question! You can certainly use both the NHEJ and HDR pathway for gene knock-out, however, HDR is more commonly used to make precise modifications (i.e. specific nucleotide changes). This is because in order to utilize HDR, you will also have to include a "repair template" containing the desired sequence as well as additional homologous sequence immediately upstream and downstream of the target. You will also have to keep in mind that the efficiency of HDR is generally low (
Got it! Thanks.
hi , i have a presentation about crispr cas9, how can i take the figures used by you
Hi Adila, thanks for leaving your comment and reaching out to us! We're glad our graphics are useful to you. Please contact info@abmGood.com with your inquiry and a list of the images you'd like to use and someone from our team will get back to you, shortly!
Will a single gRNA work for all the genes, as in the all in one vector? What are your thoughts on the use of two targets for the same gene to delete the intervening sequence?
Rachel Jesudasan Different gRNA's have different targeting efficiency, and so it is hard to predict how well a given gRNA will perform. Therefore, it is always ideal to design 2-3 different gRNA constructs against your desired gene and use them individually as well as grouped together to increase the efficiency of gene knockout. The all-in-one system provide an ideal tool for transfecting both the gRNA and the Cas9 protein with a single transfection vector which increases the transfection efficiency. This tool is especially useful when you already had success with a particular sgRNA sequence. If you will be using two or more gRNA to delete large portions of the genome it is critical to ensure that off target effects of the gRNA are minimized as it could potentially lead to unwanted off-target effects. for more information check out our latest video at: th-cam.com/video/dXPDefej0Ps/w-d-xo.html
If I want to knock out the dystrophin gene, then how would I do that as dystrophin gene is a big one.
Very soon we will be releasing our next video in this series which will walk you through the gRNA design procedures. Briefly here, we would recommend you to design your gRNA against exon 1 or 2 of the dystrophin gene; this will ensure that frame shift mutations caused by NHEJ do not lead to partially functional gene product. you could also try designing multiple gRNAs against the dystrophin gene and transfect all of them together. abm also provides these gRNA pre-designed and you could find them here: Human- www.abmgood.com/DMD-CRISPR-Lentivector-K0609305.html; Mouse - www.abmgood.com/DMD-CRISPR-Lentivector-K4939705.html; Rat - www.abmgood.com/DMD-CRISPR-Lentivector-K6920905.html
Thank you for the great video and informative Q&A. I was wondering if this technology can be used to introduce an opsin to mammalian cells?
Yes, it can! CRISPR can be used to knock-in any gene, and opsins are no exception. Here is an example of a paper that knocked in a CAG-LSL-ChR2-Tdtomato cassette: www.sciencedirect.com/science/article/abs/pii/S1534580718303277
The most common combination of CRISPR and optogenetics, however, is to create a photo-activatable Cas9. There's a good blog post about some of these photo-activatable CRISPR tools here:
blog.addgene.org/optogenetics-crispr-using-light-to-control-genome-editing.
Amasing 🎉
would a CRISPR/Cas9 work on extracted DNA?
thelongboardsling yes it will work on extracted DNA as well. The requirements will be a functional Cas9 protein and a full length gRNA (including the target sequence and the scaffold sequence). In fact there has been some recent research on this - in order to use Cas9 to perform Restriction digests and Gibson assembly (goo.gl/PQAqae)
thanks !!!!
How target site of dsDNA opens for target sequence for attachment?
Hi Zia, thanks for watching our video and leaving a comment! I believe the biophysics of the mechanism are still being heavily studied, however, what is somewhat known is that an "R loop is formed" in which the sgRNA intrudes upon the DNA double helix to form an RNA-DNA hybrid helix with the target strand and displacing the opposite nontarget strand (from this paper: goo.gl/3vB4O9).
Video's great, your voice is painful to listen to though. (or rather, the audio is)
Hi Trafalgar - this was one of our earlier videos. Our newer CRISPR videos have much better sound quality! You can get our entire playlist here: goo.gl/eaxKXK
Cure for hair loss yet?
Does any University that does these studies and experiments have a volunteer program for humans, say you have a cancer in the brain where there nothing that can be done, I wonder if this would be helpful in a cure while still alive.
+Baby Boo . This technology is very new. We are still a long way from being able to use this technology in a clinical setting. There are researchers that are working towards that goal such as Editas Medicine (editasmedicine.com/index.php).
Thank you for the answer, My pastor has a son that is 15 and he has a Very rare brain tumor[cancer] was thinking this would be great if it could be done as you say in a clinical setting, they have done all they can.
Thank you again for the answer.
+Baby Boo -You might want to tell your pastor to check out Dr. Burzynski's antineoplastons protocol, which has helped and healed many with 3rd and 4th stage brain tumors. Hope this helps...
I do know of Dr. Burzynski's and he is one of the best I believe my dad has a book/articles on or about him and we have a friend who has cancer,. I do not have any health problems but I was just wondering if any if these Universities do experiment type studies.
This may be a primer on CRISPR but it's not for the uninformed.
+Sherry Von We are sorry to hear that, if you like us to explain anything in more detail please let us know! We will either answer you here or make another video to go into that detail! also be sure to check out our knowledge base for more detailed information: goo.gl/ceUmEE
Cure HIV Crisprs Cas 9 AGT
Wow. The narrating voice in this video somehow really irritates me, the pitch of it really gets into my bones. To bad for me, because it seems like a really good video.
Hi APOLOnl, thanks for your comment! Sorry you didn't like the narration, our future videos have better sound quality. If it helps, you can always mute the video and play it with our closed-captioning turned on. We also have a comprehensive knowledge base where you can find all this information and more: goo.gl/df55ap.