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

Description of the steps required to design effective gRNA sequences and then clone those sequences into a Cas9 expression plasmid for KO experiments.

[@marazilocchi1721]

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

[@jigglypuff6297]

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!

[@plantbiotech1701]

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

[@mandarin9967]

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:)

[@ttp2210]

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.

[@uzoechisamuel]

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

[@muskangupta4542]

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

[@ahsenkonac7992]

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

[@alexdesouza8351]

The greatest explenation of CRISPR in KZbin. Congratulations

[@ManiKitchenHealthy]

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

[@roujehgashan1310]

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

[@mirkhurshid8243]

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.

[@younusbhat8019]

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

[@alicegevorgyan7046]

This was really helpful, thank you for making it!

[@gabrielletodd3314]

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

[@tramdang8924]

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

[@mariasimanovich4259]

It's just the perfect explanation. Thank you!

[@tammy1130HD]

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

[@elliem0830]

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

[@amirhosseinjafariyan9203]

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

[@mariamadelowo226]

Thanks for this very explicit video

[@aa-so4nj]

Very informative lecture with details, thanks a lot!

[@aman199kumar]

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

[@tifikocku]

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

[@cacevedo07]

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

[@moizkhan1501]

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.

[@salmaaktar7714]

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

[@yayayayapewpew]

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

[@JasonLubega]

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

[@shanidror4613]

Thank you so much! very useful and clear explanation!

[@varsharathore6457]

Thank you so much, you really explained so clear🙏

[@Forever1DFan100]

cant thank you enough for making this video!

[@ednilsonvarela1712]

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

[@mfarooqsr5607]

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

[@waqasrafique6016]

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.

[@user-xr6sk2ch2q]

Amazing explanation

[@naeemakhter5666]

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

[@hibahi9008]

Extremely helpful! Thanks a lot

[@hitkarshkushwaha2434]

I love your explanation..

[@nishattamanna2372]

More than a Perfect Video!!

[@oberschnerpfel]

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

[@Exploretheexperiece]

Very useful contents.thanks dear

[@user-ie7fx9xn9q]

Very informative video. Clear all doubts about gRNA design.

[@muhammadbilalsarwar5646]

Thanks for the wonderful informative video :) .

[@priyambiswas4902]

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

[@cristinacl3782]

Perfect explanation thank you!

[@amiteshanand9073]

Great job, Jacob!

[@rohinibhattacharya2067]

This video is amazing

[@stuart999able]

This was very helpful

[@quackat1110]

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

[@gsmaga2]

Thanks a lot for your explanation it helped a lot

[@zahrabaghban6313]

Very useful and instructive

[@anggunsausan6738]

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

[@user-lh8wc1bc6q]

awesome video!! thanks!!

[@Omidvatan]

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

[@andreadellolio805]

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

[@cr9367]

Really nice tuto thanks !

[@ARASHIop77]

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

[@AnNguyen-dp7qd]

thanks a lot, it is extremely informative !

[@9836862908]

This is outstanding

[@BestFoodAndTravelVideos]

SIMPLY PERFECT

[@yelizidilyigit5716]

thank you very much, it's very helpful!

[@aurelie8287]

This is great! Thanks!

[@mihir86100]

Really helpful. More video expected on CRISPR

[@aymankhalifa5462]

Thank you!

[@sciencehabitat3969]

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

[@laowan4064]

thanks for this tutorial.

[@mangarific1]

Thankyou so much! Please keep making engineering videos

[@trynewthings321]

Awesome Video thanks alooooooooot

[@anumjaved3702]

Superb

[@masumasultana1236]

great video indeed

[@Farhab]

Awesome 🎉

[@celcorsystems1890]

good work

[@shaliperezmendez735]

Te amo, gracias me salvaste

[@vijaytejwani461]

Bravo!

[@DeepakKumar-fo4mg]

Dear, Greetings, You are awesome

[@alvarolucci57]

Genius!

[@manikantakolasani8440]

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

[@ruilan2238]

AMAZING THANK U SO MUCH

[@mimm4332]

much love

[@diptanilbiswas5657]

Lovely

[@gorsarkisyan5614]

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.

[@enerandmackon761]

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

[@ankitavaishampayan2239]

Such a clear and informative video! I had a quick question. I am looking to knock out a gene cluster in bacteria. Would this procedure work in bacteria as well? Thank you very much 🙂

[@zarak5055]

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

[@Daniel-wd7ie]

Hi Jacob, I am glad I found your video! It is really helpful. However, I still have one question: if you have these sticky ends between gRNA and scRNA, how do they assemble correctly?

[@kiranpatil2924]

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

[@canergunaydin7145]

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

[@nahuelcesattilaluce1257]

I found your video more than useful! Thanks a lot...and I have a question! there is a way to see in what exon is located targets sequences ? I mean, not manually checking every exon in the gene... If you work with a big gene, with lot of exons, it can be pretty annoying look for your target sequence just checking sequence along.

[@Farhab]

Yes use snapgene software

[@dorfinhorg9458]

Hey this video is super helpful and well made! Do you have any supporting literature you could provide?

[@shynamiri3932]

Hi Jacob! very helpful video. Thanks. Anyone has any idea about how to make a plasmid that induces double KO for 2 genes? Or better to say, how to make a double KO cell line.

[@ryanswan4145]

A wonderful and informative video. I had one quick question, I'm looking into knockdown of a gene (INPP5D) in human genome but am wondering the main differences between the two options for homo sapiens provided by chopchop. If you could clarify the significance that would be very helpful. Thank you!

[@jakelmer1985]

Are you referring to the hg19 and hg38 versions of the human genome?

[@faizanaz7976]

Very nice video, please tell me what the efficiency column means?

[@zakiaal-hareth1508]

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!

[@himensalimizand5074]

Thank you for awesome video. Is there any website to design gRNAfor Cas3 CISPR system?

[@rajansandhu2471]

Very informative...have a little query...what that + written means ..is this target sequence found in coding strand or whether it will be targeting the coding strand..?

[@danielv563]

Dear Jacob, thank you for the nice lecture! I wonder, is there a company that if you seen the gRNA they will send you already the cloned plasmid?

[@AbuBakar-yv5qx]

i really liked your video..can you help me in designing the sgRNA for MLo-1 gene of Triticum aestivum??

[@starrani5264]

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

[@Justin-dv7ul]

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

[@uzoechisamuel]

Thanks. Do you have a Knock-In video?