Prime editing (another way to edit genes)

[@DeenyCat]

Starting at a new lab without the best background in genetics and this was soooo helpful! thank you !

[@BillKonstantakos]

Great review! Also worth mentioning that they designed sgRNAs that match only the edited strand to minimize double-strand DNA breaks (PE3b); this way nicking of the non-edited strand occurs after edited strand flap resolution.

[@TheSheekeyScienceShow]

Thank you!

[@wusam5764]

I like how you give really great, thorough introduction and that you explain the paper really clearly with many visual aids. :)

[@TheSheekeyScienceShow]

Thank you very much for your kind review!

[@shiyiyin3403]

4:36 , regarding base editing, ABE and CBE are now nickase based as of 2024, i did remember early on they were dcas9; Cas9n means N terminal fusion protein, can be confused with nCas9, nickase cas9 Great video!!!

[@treewx]

So, in Prime Editing, that Cas9 attaches to the Target Strand, but edits the Non-Target strand?

[@videofancrazy18]

Really well explained, and you posited all the questions we need to ask about this technology. Your sense of humour certainly doesn't hurt lol.

[@TheSheekeyScienceShow]

Thank you! And glad to hear you enjoy the humour 😊

[@adeelmehmood8093]

@@TheSheekeyScienceShow can be editing polycystics kidney disease which involved faulty genes their names pkd1 and pkd2 thanks

[@fazelamirvahedi9911]

That was really nice and well-explained. Thank you for sharing your knowledge and skill. Please make more videos like this.

[@inquisitivebiologist2442]

This was an actually amazing video explaining Prime editing. Thanks a lot, you are great!

[@TheSheekeyScienceShow]

Thank you very much!! 😊

[@inquisitivebiologist2442]

@@TheSheekeyScienceShow You really made this complicating thing easy to understand for me. Thanks a lot. Writing a research paper on genome editing, and I'm using Prime as the Future! Very interesting and promising!

[@chun-yinghuang8627]

Thank you for making such a great video! I’m working on gene editing and this video really helps!

[@TheSheekeyScienceShow]

Thank you very much! 😊

[@lilyz646]

Very well explained!!! I loved it thank you very much! I think you might've mislabeled two 5’ ends on the pegRNA at around 9:12 minute mark?

[@lilyz646]

The singing that started shortly after in the background scared the ever loving Jesus out of me 😂😂😂✌️

[@TheSheekeyScienceShow]

Thank you and aha, yes indeed, well spotted!

[@TheSheekeyScienceShow]

@@lilyz646 Yes, i can imagine, think it was an owl...!

[@motaherhossain4491]

Well explained. Could you please tell me which software did you use for the depiction of your video?

[@TheSheekeyScienceShow]

Thanks and 'Microsoft Whiteboard'

[@motaherhossain4491]

@@TheSheekeyScienceShow I see. Thank you.

[@BFS4321]

This is cool, thank you Eleanor, for the animated cartoon video, which helps a lot. BTW, what program you used for annotation and drawing?

[@TheSheekeyScienceShow]

Thanks! I use the apps under the "Windows Ink Workspace" that came with my laptop! Hope that helps

[@MrsSolvinden]

I'm a student currently working on an essay based on the mentioned paper and randomly ended up watching this video. This was really helpful, perfectly explained and easy to understand. Thank you for your work! Not only for the great sum up but also for the critical point of view (like in Jonathan Wilde tweets). Ps: Could anyone share what's the previous cell paper he mentions?

[@TheSheekeyScienceShow]

Thank you for your feedback!! The paper he refers to is the Nature paper describing prime editing (you'll find the link in the description!). Let me know if you meant something else! 😊

[@MrsSolvinden]

@@TheSheekeyScienceShow Thank you for the quick answer :) I meant the one he says they haven't cited. (Cell paper previous to the Nature one) I found it so I'll leave it here in case anyone else is interested! doi.org/10.1016/j.cell.2018.08.057

[@aoihana1042]

Excellent and easy to follow summary🎉

[@sanja4890]

Super good explanation! Thanks for the video and the references :)

[@shroomwave9281]

Thank you for this video. I'll give you lots of credits in my presentation!

[@kiara_l8732]

Thank you so much for making this! A quick question, is the primer binding site in the linker region between the cas9 and the reverse transcriptase?

[@Mohammadssalehi]

"Ooh exciting" lol Great job!

[@TheSheekeyScienceShow]

Haha, thank you!

[@karenmendez587]

Thank you so much for this video! it really helped. DO you have a protocol that you can share? Thanks again

[@TheSheekeyScienceShow]

Thanks! I dont have a protocol, sorry!

[@sudipaich7617]

Mam crispr can delete genetic hyperpigmentation?

[@basic48]

How was the word "Sheekey" derived?

[@shikhas425]

I cannot stop laughing at the "ooh exciting" at 8:45. Also this video is super helpful :)

[@RadwanDerbala--]

A pretty awesome video, no more no less

[@TheSheekeyScienceShow]

Thank you very much!

[@sheenasmith2195]

Very nice summary-- thanks!

[@TheSheekeyScienceShow]

Thank you! 😊

[@tgime1]

Love this explanation!

[@ٹیڑھا]

Thank you so much. This was very helpful

[@MrEstradaG]

u got yourself a new subscriber

[@yangzhou8486]

Great great video!!!

[@johanlauritsen]

Why is SSBs "inefficient", your conlusion is the direct opposite of what other researchers say, namely that DSBs are inefficient, therefore SSBs er more efficient?

[@TheSheekeyScienceShow]

The purpose of prime editing is to make gene editing safe and efficient. Using wild-type Cas9 that induces DSBs has been shown to efficient but is not safe. SSBs are much safer and use a mutated version of Cas9 that cuts one DNA strand. SSBs are quickly repaired by the cell which does not provide sufficient time & interaction for the integration of the sgRNA/Cas9/repair template complex, hence very low efficiency - this was the rationale behind the whole paper and design of prime editing. I have personally tested and directly compared the editing efficiencies of WT-Cas9 (DSBs) and H840A-Cas9 (SSB) and in my set up WT efficiency was ~20% whilst H840A was

[@IsaacOLEG]

OSX or WINDOWS ?

[@TheSheekeyScienceShow]

Windows

[@bretcaro8478]

Great explanation!

[@brickmortar6965]

thx. Very helpful overview.

[@zeminren6020]

Great review

[@TheSheekeyScienceShow]

Thank you! 😊

[@ahmetkara2330]

wow! really helpful

[@TheSheekeyScienceShow]

Thanks! 😊

[@ShyCataclysm]

good job

[@TheSheekeyScienceShow]

Thank you! 😊

[@gregsmith6756]

This is too complicated. I will stick with near PAM-less CRISPR, using HDR donor repair templates.