Designing gRNA Oligos to Clone into Cas9 Expression Plasmids for KO Experiments

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  • Опубліковано 24 лис 2024

КОМЕНТАРІ • 151

  • @marazilocchi1721
    @marazilocchi1721 3 роки тому +49

    I read 200 papers to understand CRISPR knockout and it was almost impossible. This is the best explanation ever. Simple and efficient. Thanks!

  • @plantbiotech1701
    @plantbiotech1701 7 місяців тому +3

    such a great videos i read so many papers but nobody explained this clearly

  • @MJ_00-q1y
    @MJ_00-q1y 4 місяці тому +4

    I can’t believe someone can explain this so simply while being detailed! Thanks Elmer! 👍

  • @jigglypuff6297
    @jigglypuff6297 3 роки тому +17

    I am usually skeptical of long videos, but this one explained everything I needed to know well and did not waste a single second. Thanks!

  • @mandarin9967
    @mandarin9967 2 роки тому +1

    Thank you soooo much! This video is extremely helpful. Should mention that you have a nice voice and pronunciation, it's so important for a non-native speaker:)

  • @alexdesouza8351
    @alexdesouza8351 Рік тому

    The greatest explenation of CRISPR in UA-cam. Congratulations

  • @uzoechisamuel
    @uzoechisamuel 3 роки тому

    I paid fees to people that confused me. In less than 30mins you explained everything. Thanks so much.

  • @muskangupta4542
    @muskangupta4542 4 роки тому +12

    Probably the best video..... your efforts are really appreciated:-)

  • @ahsenkonac7992
    @ahsenkonac7992 4 роки тому +11

    this video much more informative and clear than classical procedures, especially the chopchop part is very helpful. . Thanks a lot for sharing.

  • @ttp2210
    @ttp2210 11 місяців тому

    Thank you so much. The way you explain by breaking everything down makes it so easy to understand. I hope you make more videos like this. Also, your way of articulation and slides are amazing and never bores me out.

  • @younusbhat8019
    @younusbhat8019 2 роки тому

    Thanks a lot Jacob... .. . Even few of my Postdoc Colleagues were not able to make me understand it like the way you did... .. . Really appreciate it a lot

  • @ednilsonvarela1712
    @ednilsonvarela1712 2 роки тому

    Congratulations Jacob Elmer, I really like it. Super didactic. Surely one of the best video.

  • @DrKGJainAIIMSNewDelhi
    @DrKGJainAIIMSNewDelhi 10 місяців тому

    Very informative video. Clear all doubts about gRNA design.

  • @JasonLubega
    @JasonLubega Рік тому

    Grateful for this video. It has been helpful. You are a good teacher.

  • @mirkhurshid8243
    @mirkhurshid8243 Рік тому

    i have never done CRISPER, after watching this great master lecture i am confident that I can do it very well in lab. very detailed practical steps.

  • @cacevedo07
    @cacevedo07 2 роки тому

    Best tutorial on this topic, I've ever watched. You have another subscriber here. Many thanks!

  • @amalia4693
    @amalia4693 2 місяці тому

    Excelente trabajo explicando ❤ Muchísimas gracias!

  • @yayayayapewpew
    @yayayayapewpew 10 місяців тому

    So beginner friendly and helpful. Thank you so much! ♥️

  • @amirhosseinjafariyan9203
    @amirhosseinjafariyan9203 Рік тому

    This video was amazing, Jacob! thank you so much for sharing it!

  • @elliem0830
    @elliem0830 Рік тому

    OMG!!!! FINALLY, I GOT IT! This video is the best ever Jacob:) Thank you

  • @aman199kumar
    @aman199kumar 2 роки тому +2

    Very informative especially for the beginners. Thank you for making this video!

  • @moizkhan1501
    @moizkhan1501 Рік тому

    Thank you. Please make more videos on techniques used in the field of Biotechnology. The way you explain procedures and processes is very easy to understand and follow. You, sir, are amazing.

  • @ManiKitchenHealthy
    @ManiKitchenHealthy 9 місяців тому

    very good video. Easy to understand such a difficult approach.

  • @tramdang8924
    @tramdang8924 3 роки тому

    This is the best video I have found on this topic. Thank you.

  • @naeemakhter5666
    @naeemakhter5666 8 місяців тому

    Amazing video easy to understand and you have a really nice way of explaining ! Keep up the good work

  • @nishattamanna2372
    @nishattamanna2372 3 роки тому +1

    More than a Perfect Video!!

  • @tammy1130HD
    @tammy1130HD Рік тому

    thank you for making this video in such clear and practical way!

  • @sciencehabitat3969
    @sciencehabitat3969 3 роки тому

    My first word after watching this video was WoW. Not usual for me.

  • @mariasimanovich4259
    @mariasimanovich4259 2 роки тому

    It's just the perfect explanation. Thank you!

  • @yourlifeonpower
    @yourlifeonpower 4 місяці тому

    This is a fantastic explanation; thank you so much!

  • @quackat1110
    @quackat1110 2 роки тому +1

    Utterly resourceful, definitely deserve more views and subscriptions!!
    Btw, chopchop is constantly in use, priority 6-9 is common..

  • @roujehgashan1310
    @roujehgashan1310 4 роки тому +1

    Thank you so much for the simplicity and clarity of the instructions

  • @priyambiswas4902
    @priyambiswas4902 4 роки тому

    So nice presentation... Very informative & easily digestible material.. Much useful than other Videos.. Thanks a lot

  • @oberschnerpfel
    @oberschnerpfel 4 роки тому +1

    Great video man, my professor just isn't all there and no one in our course understood how to design a gRNA :/ With your Video I just get it now. Thanks for the clarity and especially the explanation how the algorithm works and how you could do it yourself. Such a great help thx again

  • @Omidvatan
    @Omidvatan 4 роки тому +3

    I have been looking for a good source for gRNA design for several weeks. Only with this video, I could find the answer to almost all of my questions. Thanks a lot, Jacob. Great job. Would you please add a quick video on how to design gRNA for CRISPR/dCas9 system for gene expression regulation? Thanks

  • @gabrielletodd3314
    @gabrielletodd3314 2 роки тому

    Thanks so much! This was incredibly helpful and I appreciate all of the useful tidbits!

  • @tifikocku
    @tifikocku 4 роки тому

    Dude, this is thy best CRISPR video on the net! tnx

  • @LOTUS-pr3fl
    @LOTUS-pr3fl 2 місяці тому

    Amazing explanation! Please make more video related this topics! it's better than reading 1000 paper!
    Recently I am coming UA-cam to see your video!
    already subscribed and clicked bell icon

  • @Exploretheexperiece
    @Exploretheexperiece 2 роки тому

    Very useful contents.thanks dear

  • @muhammadbilalsarwar5646
    @muhammadbilalsarwar5646 2 роки тому

    Thanks for the wonderful informative video :) .

  • @enerandmackon761
    @enerandmackon761 3 роки тому +1

    the best video! thank you, please can you continue this video for the next step of CRISPR/cas9?

  • @ARASHIop77
    @ARASHIop77 2 роки тому +1

    Thanks jacob. I loved how finely you explained everything. I was wondering if you make a video on crisprs mediated repair outcomes.

  • @mfarooqsr5607
    @mfarooqsr5607 4 роки тому

    Sir, very well informative lecture for me I loved the way you explained.

  • @mariamadelowo226
    @mariamadelowo226 4 роки тому

    Thanks for this very explicit video

  • @תהילהבןזיו
    @תהילהבןזיו Рік тому

    Amazing explanation

  • @aa-so4nj
    @aa-so4nj 3 роки тому

    Very informative lecture with details, thanks a lot!

  • @Forever1DFan100
    @Forever1DFan100 3 роки тому

    cant thank you enough for making this video!

  • @Farhab
    @Farhab Рік тому

    Awesome 🎉

  • @mangarific1
    @mangarific1 3 роки тому

    Thankyou so much! Please keep making engineering videos

  • @cristinacl3782
    @cristinacl3782 2 роки тому

    Perfect explanation thank you!

  • @BestFoodAndTravelVideos
    @BestFoodAndTravelVideos 7 місяців тому

    SIMPLY PERFECT

  • @mihir86100
    @mihir86100 3 роки тому

    Really helpful. More video expected on CRISPR

  • @salmaaktar7714
    @salmaaktar7714 4 роки тому

    thank u for simple but nice explanation, it helps me a lot

  • @alicegevorgyan7046
    @alicegevorgyan7046 4 роки тому +2

    This was really helpful, thank you for making it!

  • @hitkarshkushwaha2434
    @hitkarshkushwaha2434 4 роки тому

    I love your explanation..

  • @zahravahramian
    @zahravahramian 2 роки тому

    Very useful and instructive

  • @waqasrafique6016
    @waqasrafique6016 3 роки тому

    Comprehensive details, the Best method of explanation. I am gonna subscribe and like your channel to support your generous efforts. Thanks btw for the time and effort you put into this video.

  • @manikantakolasani8440
    @manikantakolasani8440 4 роки тому +1

    sir u are amazing
    I really loved the way u explained
    u made my job much easier
    plz make an video how do I clone a dna segment in to a vector in detail

  • @hitkarshkushwaha1727
    @hitkarshkushwaha1727 5 місяців тому

    Very nice dear sir

  • @gsmaga2
    @gsmaga2 3 роки тому

    Thanks a lot for your explanation it helped a lot

  • @hibahi9008
    @hibahi9008 3 роки тому

    Extremely helpful! Thanks a lot

  • @rohinibhattacharya2067
    @rohinibhattacharya2067 3 роки тому

    This video is amazing

  • @shanidror4613
    @shanidror4613 4 роки тому

    Thank you so much! very useful and clear explanation!

  • @varsharathore6457
    @varsharathore6457 3 роки тому

    Thank you so much, you really explained so clear🙏

  • @AnNguyen-dp7qd
    @AnNguyen-dp7qd 2 роки тому

    thanks a lot, it is extremely informative !

  • @9836862908
    @9836862908 3 роки тому

    This is outstanding

  • @anggunsausan6738
    @anggunsausan6738 4 роки тому

    Oh my God, you save my life. Thanks a lot!

  • @andreadellolio805
    @andreadellolio805 3 роки тому

    Thanks thanks thanks for the help! You're a legend!

  • @laowan4064
    @laowan4064 3 роки тому

    thanks for this tutorial.

  • @정현석-n2s
    @정현석-n2s 2 роки тому

    awesome video!! thanks!!

  • @stuart999able
    @stuart999able Рік тому

    This was very helpful

  • @kiranpatil2924
    @kiranpatil2924 2 роки тому

    Thank you,, Please make a video on Analysis of ON and OFF target from NGS seq.. Cheers

  • @yelizidilyigit5716
    @yelizidilyigit5716 3 роки тому

    thank you very much, it's very helpful!

  • @aurelie8287
    @aurelie8287 4 роки тому

    This is great! Thanks!

  • @masumasultana1236
    @masumasultana1236 3 роки тому

    great video indeed

  • @trynewthings321
    @trynewthings321 3 роки тому

    Awesome Video thanks alooooooooot

  • @cr9367
    @cr9367 5 років тому

    Really nice tuto thanks !

  • @ramwungshimrahh3787
    @ramwungshimrahh3787 4 роки тому

    Thank you!

  • @mimm4332
    @mimm4332 Рік тому

    much love

  • @alvarolucci57
    @alvarolucci57 2 роки тому

    Genius!

  • @vijaytejwani461
    @vijaytejwani461 3 роки тому

    Bravo!

  • @anumjaved3702
    @anumjaved3702 3 роки тому

    Superb

  • @komal9405
    @komal9405 2 місяці тому

    Thank you so much for the excellent video! I have a few questions about guide RNA (gRNA) design that I hope you can help with. I'm working on a gene with three exons and two introns. The first two exons are in the 5' UTR, and the third exon is partially in the 3' UTR. When I try to design gRNAs for exon 1 and exon 2, I struggle to find optimal matches. Even when one meets criteria like GC content and self-complementarity, it still has mismatches, some of which are in the exons or CDS regions. Following your method from the video, I input the gene ID and found the exon blue blocks and intron lines. However, I couldn't identify suitable gRNAs for exon 1 and exon 2, while exon 3 displays many potential gRNAs. Since I’m still new to gRNA design, I'd really appreciate your guidance. Should I focus on designing separate gRNAs for exon 1 and exon 2, or should I concentrate on exon 3, given the better matches? Your advice would be incredibly helpful!

  • @aminasaleem7428
    @aminasaleem7428 Рік тому

    Hi, your video explained all the aspects clearly. I just want to know about the strand option there are"'+" and "_" signs in that column. Which one we should select? I am confused here.

    • @Farhab
      @Farhab Рік тому

      Can select anyone. Both are right.

  • @ruilan2238
    @ruilan2238 3 роки тому

    AMAZING THANK U SO MUCH

  • @celcorsystems1890
    @celcorsystems1890 Рік тому

    good work

  • @diptanilbiswas5657
    @diptanilbiswas5657 3 роки тому

    Lovely

  • @DeepakKumar-fo4mg
    @DeepakKumar-fo4mg 4 роки тому

    Dear, Greetings, You are awesome

  • @Kenny_Log-in
    @Kenny_Log-in Рік тому

    I love your content, and absolutely appreciate the offer to check our gRNAs. I'm sure there's limits to how much time you can spare, but when it's most important, I will take you up on that offer.

  • @gorsarkisyan5614
    @gorsarkisyan5614 2 роки тому

    Great video, thanks for taking the time to explain how it works, this is great help. I have one question, how should I proceed if I want to excise a potion of an intron and remove it completely? Thanks in advance.

  • @canergunaydin7145
    @canergunaydin7145 2 роки тому

    Informative! Can you also make a video about knock-in design ? Thanks.

  • @shaliperezmendez735
    @shaliperezmendez735 2 роки тому

    Te amo, gracias me salvaste

  • @zarak5055
    @zarak5055 2 роки тому

    That was a great video. Thank you very much. How about if we want to Knock in a gene?!

  • @uzoechisamuel
    @uzoechisamuel 2 роки тому

    Thanks. Do you have a Knock-In video?

  • @alefeamorim727
    @alefeamorim727 4 роки тому

    Hi Jacob,
    I am currently undertaking a CRISPR.Cas9 experiment with Wheat for my masters degree and Chop-CHop isn't working for me neither Synthego - keep getting erros. You said in the video you would be happy to help , so just giving a long shot here in case you read this msg!

  • @zakiaal-hareth1508
    @zakiaal-hareth1508 2 роки тому

    Thank you, the video is a great help; but what about the scaffold RNA? is it included in the plasmid in addition to Cas9 encoding gene?
    In this case, we do not design the plasmid, we purchase it from some company!

  • @kalpitakaran
    @kalpitakaran 7 місяців тому

    Excellent explanation.

  • @jedn6488
    @jedn6488 Рік тому

    Hi why are there two complementary strands? I thought crispr was single guide strand. How will these attach to the genomic dna if they are attached to themselves?

  • @ayodejitolu2652
    @ayodejitolu2652 2 місяці тому

    i have just one question. How will I link Cas 9 protein, my gRNA, and those promoters to plasmid. if you can do a video on that, i will really appreciate

  • @starrani5264
    @starrani5264 4 роки тому

    thanks a lot,... so well explained , could you please explain more about NGG sequence ? thanks

    • @Justin-dv7ul
      @Justin-dv7ul 3 роки тому

      basically its the PAM sequence and I think its used as a way to identify the difference between the gRNA and the sequence