theCrux
theCrux
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dCas9: CRISPRi CRISPRa CRISPR-ON CRISPR-OFF (Beyond Cas9 Genome Editing)
References/Resources: www.patreon.com/the_Crux
All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html
What is CRISPRi (CRISPR interference)? What is CRISPRa (CRISPR activation)? What is a dCas9 and how to use dCas9 for CRISPRi and CRISPRa experiments? dCas9 can be fused with effector domains to regulate gene expression for functional studies. In addition, you can extend the CRISPRi/CRISPRa based tools to build permanent epigenetic modifications using CRISPR-ON and CRISPR-OFF. In this video, I discuss the details of dCas9 based genetic manipulations (and multiple iterations/generations of these tools) used to repress and activate genes and go beyond genome editing using CRISPR/Cas systems.
00:00 dCas9
00:52 CRISPRi (Gen 1)
02:21 CRISPRa (Gen 1)
03:36 CRISPRa (Gen 2)
06:38 Challenges
07:41 CRISPRi/a (Gen 3)
10:32 CRISPR-ON
11:38 CRISPR-OFF
CRISPR-Cas9 in genome editing: th-cam.com/video/7ESZTE6rjLI/w-d-xo.html
Epigenetic Replication: th-cam.com/video/HB4IAiYqdzA/w-d-xo.html
References/Resources: www.patreon.com/the_Crux
*Must watch playlists*
All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html
All videos on DNA Replication: th-cam.com/play/PL0Ymnd-zt4IgLefDaeqicJbCBnbLqnDJ7.html
All videos on Transcription: th-cam.com/play/PL0Ymnd-zt4Ij2VcAOHNUweftSElVsc-MX.html
All videos on Translation: th-cam.com/play/PL0Ymnd-zt4IjAAAMlPLDIXbs692Yp14xa.html
Consider supporting the growth of this channel by becoming a Patron: www.patreon.com/the_Crux
I appreciate your support :)
มุมมอง: 701

วีดีโอ

CRISPR KO vs CRISPR KI experiment using CRISPR-Cas9
มุมมอง 506หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html How to design a CRISPR experiment? How to perform surveyor assay to check gRNA targeting efficiency? How to perform a CRISPR knock-out experiment? How to perform a CRISPR knock-in experiment? What is the difference between the steps to perform a crispr deleti...
How to design and express CRISPR guide RNA [pX330 plasmid, Tutorial]
มุมมอง 862หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html What is a typical CRISPR Cas9 protocol? What is the workflow of a typical CRISPR protocol? How to design CRISPR Cas9 guide RNA for a CRISPR experiment and clone crispr guides in a plasmid (pX330)? In this video, I discuss a typical CRISPR protocol workflow an...
Genome Editing with CRISPR-Cas9
มุมมอง 6Kหลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html What is CRISPR-Cas9 technology? How does CRISPR-Cas9 genome editing work? What is the mechanism of genome engineering with CRISPR-Cas9? What is a PAM sequence? Where is the PAM sequence location and what is its function? What is the protospacer adjacent motif...
CRISPR-Cas9-mediated Adaptive Immunity Mechanism in Bacteria [3 Steps of CRISPR-Cas system]
มุมมอง 1.5K2 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html What is CRISPR-Cas9? How does CRISPR-Cas9 work in its natural context? How does CRISPR-Cas9 protect bacteria against pathogens by providing adaptive immunity? What are the different types of CRISPR systems? CRISPR-Cas9 is a Type II-A CRISPR system. In general...
A Brief History of Genome Editing
มุมมอง 5382 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html This video briefly discusses the history of genome editing and how historical events have led to the current state of the art. The video discusses parallel timelines that have interwoven to result in modern methods today. It all of course rests on the discove...
Protein Expression Vectors - tet-OFF and tet-ON (tTA/rtTA + Tet based gene regulation) [Part 5]
มุมมอง 9362 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html What are the types of tet expression systems? What is a tet-ON and tet-OFF expression systems? How do tetracycline inducible expression systems work? This video discusses the regulation of protein expression using a tetracycline inducible or repressible based...
Protein Expression Vectors (pET vector) - Induction of Protein Expression (IPTG + T7 Pol) [Part 4]
มุมมอง 1.5K2 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html What is a pET vector? What is a pET expression system? How do pET vector expression systems work? What are inducible expression systems and how do pET vector systems work? What is the function of T7 RNA polymerase in pET vectors and how does T7 RNA polymerase...
Protein Expression Vectors - Examples (Fusion Proteins, Insulin, and Cas9) [Part 3]
มุมมอง 5533 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html In this video, we explore some common protein expression vector designs and how they are constructed. These are helpful examples and applications of protein expression vectors to intuitively understand the protein expression vector construction. We discuss 5 ...
Protein Expression Vectors - Template, Codon Bias, Affinity Tags and Epitopes [Part 2]
มุมมอง 5143 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html In this video, we explore how to obtain templates for protein expression vectors depending on the gene structure (intron and exon language). Prokaryotic genes are intron-less, whereas eukaryotic genes often have introns. Obtaining template for protein express...
Protein Expression Vectors - Expression Host and Recombinant Proteins [Part 1]
มุมมอง 8773 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html Protein Expression vectors are used to make protein from the template DNA. In this video, we discuss some of the common applications of protein expression vectors and the complications involved with protein expression when changing the host i.e. heterologous ...
RNA Expression Vectors - Design, Applications, and Examples (In vitro Transcription)
มุมมอง 4513 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html RNA Expression vectors (different from protein expression vectors) are used for the sole purpose of making RNA from a template DNA. In this video, we discuss some of the common applications of RNA expression vectors and their rules/principles of design. Final...
Golden Gate Cloning or assembly - Type IIS (BsaI) restriction enzyme based cloning
มุมมอง 2.7K6 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html Golden Gate Cloning or Golden gate assembly is a restriction enzyme based cloning that uses type IIS restriction enzymes. It is a powerful way to clone large numbers of inserts in a single reaction without the need of using multiple enzymes. This video discus...
Gateway Cloning: Simple and Multisite Gateway Cloning - LR and BP Cloning
มุมมอง 2.6K7 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html Gateway Cloning is a popular recombination based cloning method. It does not rely on restriction enzymes but instead uses integrases to recombine att sites between a final and donor vector. This causes an exchange of DNA segments that inserts the desired DNA ...
TOPO Cloning - TOPO-Blunt, TOPO-TA, TOPO-directional
มุมมอง 4.6K11 หลายเดือนก่อน
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: th-cam.com/play/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf.html TOPO Cloning (Topoisomerase-based cloning) is a commonly used method to clone PCR amplicons. TOPO Cloning bypasses the need of restriction enzymes and ligases. In this video, we discuss the three main types of TOPO-cloning systems - TOPO Blunt, TOPO TA, and T...
Cloning with Restriction Enzymes (5 Levels) - Traditional Molecular Cloning
มุมมอง 1.6Kปีที่แล้ว
Cloning with Restriction Enzymes (5 Levels) - Traditional Molecular Cloning
Plant cloning vectors - Disarmed Ti plasmid, Cointegrate vectors, Binary vectors - T DNA transfer
มุมมอง 6Kปีที่แล้ว
Plant cloning vectors - Disarmed Ti plasmid, Cointegrate vectors, Binary vectors - T DNA transfer
Yeast cloning vectors - YIp, YEp, YRp, YCp, YACs - Shuttle plasmid vectors
มุมมอง 6Kปีที่แล้ว
Yeast cloning vectors - YIp, YEp, YRp, YCp, YACs - Shuttle plasmid vectors
P1 Phage and PAC cloning vector - Pac sites, Cre-loxP, and SacB lethality
มุมมอง 2.1Kปีที่แล้ว
P1 Phage and PAC cloning vector - Pac sites, Cre-loxP, and SacB lethality
F plasmid, BACs, Fosmid cloning vector - Fosmids are small BACs (Bacterial Artificial Chromosomes)
มุมมอง 1.7Kปีที่แล้ว
F plasmid, BACs, Fosmid cloning vector - Fosmids are small BACs (Bacterial Artificial Chromosomes)
Lambda Phage vector, Cosmid cloning vector - spi phenotype and in vitro packaged lambda virus
มุมมอง 7Kปีที่แล้ว
Lambda Phage vector, Cosmid cloning vector - spi phenotype and in vitro packaged lambda virus
Plasmid and Phagemid cloning vector - Selectable vs. Screening marker
มุมมอง 4.4Kปีที่แล้ว
Plasmid and Phagemid cloning vector - Selectable vs. Screening marker
Dam and Dcm methylation (restriction independent methyl-transferases) affects restriction enzymes
มุมมอง 2Kปีที่แล้ว
Dam and Dcm methylation (restriction independent methyl-transferases) affects restriction enzymes
Restriction Enzymes (Endonucleases) in Molecular Cloning
มุมมอง 2Kปีที่แล้ว
Restriction Enzymes (Endonucleases) in Molecular Cloning
Genetic Engineering - Classic vs Modern methods
มุมมอง 3.7Kปีที่แล้ว
Genetic Engineering - Classic vs Modern methods
RNA Polymerase III Transcription - Promoters, TFs, Initiation, Elongation, and Termination steps
มุมมอง 4.4Kปีที่แล้ว
RNA Polymerase III Transcription - Promoters, TFs, Initiation, Elongation, and Termination steps
Prokaryotic Transcription Factors in Elongation and Termination (GreA, GreB, Mfd, NusA)
มุมมอง 2.6Kปีที่แล้ว
Prokaryotic Transcription Factors in Elongation and Termination (GreA, GreB, Mfd, NusA)
Rolling Circle DNA Replication and Amplification - Plasmids and Bacteriophages (M13 + PhiX174)
มุมมอง 12Kปีที่แล้ว
Rolling Circle DNA Replication and Amplification - Plasmids and Bacteriophages (M13 PhiX174)
Mitochondrial DNA (mtDNA) Replication - D loop Replication (deep-dive video)
มุมมอง 14Kปีที่แล้ว
Mitochondrial DNA (mtDNA) Replication - D loop Replication (deep-dive video)
Telomere DNA Replication - Chromosome end replication problem in eukaryotes
มุมมอง 6Kปีที่แล้ว
Telomere DNA Replication - Chromosome end replication problem in eukaryotes

ความคิดเห็น

  • @lovekenmaforlife3406
    @lovekenmaforlife3406 2 วันที่ผ่านมา

    thank you!!!

  • @aditikaushal1611
    @aditikaushal1611 3 วันที่ผ่านมา

    The explanation was so good . Thanks for the video 🤍

  • @aflahshadilmp1818
    @aflahshadilmp1818 8 วันที่ผ่านมา

    Amazing lecture sir. keep up the good work.🖤🖤

  • @chandandeosthali6625
    @chandandeosthali6625 8 วันที่ผ่านมา

    Amazing board work and nice explanation!!! Thank you...

  • @arnabghosh676
    @arnabghosh676 8 วันที่ผ่านมา

    Man..I love your videos...can you suggest any books for this topic??...It would be great if you suggest books in your descriptions.

  • @deepikaosmania
    @deepikaosmania 9 วันที่ผ่านมา

    Sir, I am a great fan of your videos . I have request. Can you please make a video on rRNA processing including ETS and ITS removal and other details

  • @sergiumitrea6437
    @sergiumitrea6437 10 วันที่ผ่านมา

    Hello from România, what i can do if i remove 300 pear tree with crown gall? How much years i cant plant in that area? In romania the enginner say 5 years to no plant nothing in that areea . Thanks.

  • @lovekenmaforlife3406
    @lovekenmaforlife3406 10 วันที่ผ่านมา

    yooo tysm!!!!!!

  • @jimmiosias3555
    @jimmiosias3555 11 วันที่ผ่านมา

    thank you ❤

  • @AmnaYaseen-jq8rw
    @AmnaYaseen-jq8rw 12 วันที่ผ่านมา

    great well explained

  • @ZnZn-qo9ol
    @ZnZn-qo9ol 12 วันที่ผ่านมา

    Big thanks from China!!!! I can still get the other techniques by reading review articles, but struggled to understand this one on my own. Real life saver!!!

  • @genisessay
    @genisessay 12 วันที่ผ่านมา

    Bro i am serious, you are literally saving my life. Why stop, please keep going. The best explanation I've ever seen and its in great detail, its perfect...

  • @marcellasidella8419
    @marcellasidella8419 13 วันที่ผ่านมา

    Sir, I am not clear how loop formation can solve the problem of incompatibility between primase and helicase. Could you please help me? 17:13

  • @kkmarskk
    @kkmarskk 14 วันที่ผ่านมา

    Crystal clear explanation, very helpful for exams

  • @mourifat8659
    @mourifat8659 14 วันที่ผ่านมา

    Is there any lab in your knowledge which offers PhD position in core genetic engineering ?

  • @deonyz
    @deonyz 15 วันที่ผ่านมา

    Very educative description! Thank you!

  • @everythingisconnected1988
    @everythingisconnected1988 16 วันที่ผ่านมา

    Amazingly clear video, thank you! Just a small remark for you: it's 'foreign', not 'foriegn' :)

  • @marygraceconcepcioncantale1640
    @marygraceconcepcioncantale1640 17 วันที่ผ่านมา

    I love how you discuss these lessons, I am a bit of a visual learner and your videos help me a lot <3

  • @umaananth3602
    @umaananth3602 18 วันที่ผ่านมา

    Excellent future effectors .........vpr's of 3 proteins called RTA - or synergistic activators

  • @AbidHasan007
    @AbidHasan007 18 วันที่ผ่านมา

    Lots of thanks. Awesome! You made my day. Just a few minutes ago, I understood the whole topic and was ready for my exam .

  • @GromGoVrom
    @GromGoVrom 18 วันที่ผ่านมา

    I appreciate your ability to take all the information about this subject and package it into a thorough lecture. Thank you very much.

  • @antoineroquentin2852
    @antoineroquentin2852 19 วันที่ผ่านมา

    Yes bro thank you so much! I’m also subbed on your patreon!

  • @theCrux
    @theCrux 19 วันที่ผ่านมา

    Epigenetic Replication: th-cam.com/video/HB4IAiYqdzA/w-d-xo.html CRISPR-Cas9 in Genome Editing: th-cam.com/video/7ESZTE6rjLI/w-d-xo.html CRISPR-KI CRISPR-KO: th-cam.com/video/f0Q6JKu8Xjk/w-d-xo.html

  • @deepthijojikarivelil5054
    @deepthijojikarivelil5054 22 วันที่ผ่านมา

    The Primase and helicase incompatibility was something that I struggled to understand! Thanks a lot for this video!!

  • @samueldl1212
    @samueldl1212 23 วันที่ผ่านมา

    Awesome

  • @samueldl1212
    @samueldl1212 23 วันที่ผ่านมา

    How tf only 800 views 😭😭😭

  • @clarat.3740
    @clarat.3740 25 วันที่ผ่านมา

    thank you so much this is a lifesaver. i've been reading the papers on golden gate and your explanation is so helpful. seeing everything drawn out makes it much easier to understand!

  • @TouibahNaseem
    @TouibahNaseem 26 วันที่ผ่านมา

    I read a textbook for this topic and got massive headache. Then afterwards watched your video understood everything thank you. It was really helpful.

  • @bhaswatigoswami244
    @bhaswatigoswami244 26 วันที่ผ่านมา

    Is this explanation masters level?

  • @diyabhatt9127
    @diyabhatt9127 27 วันที่ผ่านมา

    Thankyou for the explanation

  • @Wobbleb
    @Wobbleb หลายเดือนก่อน

    Respect 🫡

  • @SuryaprakashTripathy
    @SuryaprakashTripathy หลายเดือนก่อน

    Very nice explanation!

  • @georgeseese
    @georgeseese หลายเดือนก่อน

    Where is there an explanation for how this gene has a unique identification?

  • @earth2k66
    @earth2k66 หลายเดือนก่อน

    Your voice is heavenly, and your videos are educational ASMR. Every complex detail is organised to perfection. I would rather watch your playlists for hours than read a book or articles for 15mins 🙏

  • @earth2k66
    @earth2k66 หลายเดือนก่อน

    0:06 "Bacteria face routine harassment from Bacteriophages" 😂👌

  • @maxMax-q2c
    @maxMax-q2c หลายเดือนก่อน

    Thank you for the great videos.

  • @marlentoktomamatov5868
    @marlentoktomamatov5868 หลายเดือนก่อน

    bro this is gold

  • @marlentoktomamatov5868
    @marlentoktomamatov5868 หลายเดือนก่อน

    базара нет, шикарный видос

  • @EdT.-xt6yv
    @EdT.-xt6yv หลายเดือนก่อน

    Accelerationism/transhumanism/anthrophocene

  • @Middlebenchers-p4e
    @Middlebenchers-p4e หลายเดือนก่อน

    Sir please make a video on lytic and lysogenic induction in bacteriophage

  • @markambrozyak79
    @markambrozyak79 หลายเดือนก่อน

    😏😏😏

  • @kianjazz
    @kianjazz หลายเดือนก่อน

    YAWA WALA NAKO KASABOT

  • @theCrux
    @theCrux หลายเดือนก่อน

    CRISPR-Cas9 in Genome editing: th-cam.com/video/7ESZTE6rjLI/w-d-xo.html gRNA Design using CRISPOR and CHOPCHOP: th-cam.com/video/r4puDgc9rBs/w-d-xo.html CRISPRi vs CRISPRa: th-cam.com/video/bRlQcIAtThk/w-d-xo.html

  • @fabian_7264
    @fabian_7264 หลายเดือนก่อน

    Is it beacuse the glucose from lactose is far from the membrane where adenyla cyclase is so it can enter glicolisis before reaching the membrane therefore the inhibicion is low, in the other hand when glucose enters thru membrane proteins it is very close to adenilate cyclase

  • @christianecampos5457
    @christianecampos5457 หลายเดือนก่อน

    Very well made and extremely detailed video! It helped me understand the topic better so I could explain it in a test. Thank you very much, greetings from Brazil!

  • @user-rn3zu4no8q
    @user-rn3zu4no8q หลายเดือนก่อน

    Absolutely love your lectures, i have been reading genes by lewins along with the videos and i have to sometimes yours are even more detail oriented than them.

  • @biopharmmarkets2115
    @biopharmmarkets2115 หลายเดือนก่อน

    I'm getting into yeast genetic engineering for a project I'm working on during my Ph.D. This is an extremely useful video on a topic that is not well discussed outside of review articles

  • @theCrux
    @theCrux หลายเดือนก่อน

    CRISPR-Cas9 in Genome Editing : th-cam.com/video/7ESZTE6rjLI/w-d-xo.html CRISPR KO vs CRISPR KI: th-cam.com/video/f0Q6JKu8Xjk/w-d-xo.html CRISPRi vs CRISPRa: th-cam.com/video/bRlQcIAtThk/w-d-xo.html

  • @HarmeetSingh-jo3sg
    @HarmeetSingh-jo3sg หลายเดือนก่อน

    Superb vid!

  • @shanu6633
    @shanu6633 หลายเดือนก่อน

    Thank you dear sir. Sir can u provide notes of your every lecture please that is very helpful for me.🙏