Transcriptional Control

Figure 1: Localization of Transcriptional Effector Molecules to a genetic region of interest by attachment to a dCas9 (non-catalytically active Cas9) and guide RNA (original figure)

Mutation of the catalytic amino acids of the RuvC and HNH nuclease domains results in the creation of a catalytically inactive Cas9 protein, or dCas9, that still retains its ability to very specifically and efficiently bind double-stranded DNA sequences (Gasiunas et al. 2012).   The dCas9 and guide RNA complex can be further modified to localize a transcriptional effector molecule to a gene or region of interest for very nuanced genome modifications (Mali et al. Sept 2013).  For genome engineering purposes, this effector could be a DNA methylase to suppress the transcription of certain genes, or a histone acetylase to expose DNA for activation of transcription (Mali et al. Sept 2013; Gilbert et al. 2013).  Fusion of dCas9 to transcription factors, such as the RNA polymerase omega factor for bacteria (Bickard et al. 2013), or human heat shock factor 1 for humans (Konermann et al. 2014), results in activation of the adjacent gene. Fusion of transcriptional repression domains such as a Krüppel associated box (KRAB) domain to dCas9 results in the formation of a highly effective transcriptional repressor (Mali et al. Sept 2013).  Thus, fusion of Transcriptional Effector molecules to a dCas9 protein with guide RNA results in the creation of a highly nuanced tool for modifying expression levels of almost any gene.

Fusion of molecules to dCas9 is not just an engineering tool, it can also be used for more effective studying of specific genes.  Fusion of an activator or repressor to dCas9 in order to target specific sequences can help identify the functions of new genes (Konnermann et al. 2014).  Fusion of fluorescent proteins or molecules, such as quantum dots, to dCas9 can be used to locate the regions of DNA containing these specific sequences (Sternberg et al. 2014).

competitive inhibition transcription

Figure 2: Competitive inhibition of Endogenous DNA Binding Factors by dCas9 (non-catalytically active Cas9) and guide RNA (original figure)

The catalytically inactive dCas9 can also be used to competitively bind regions of DNA, to prevent the binding of endogenous DNA-binding factors (Mali et al. Sept 2014).  This includes RNA polymerase, and dCas9 has been demonstrated to very effectively inhibit transcription when bound to either the promoter sequence, to prevent RNA polymerase binding in the first place, or along the transcribed gene, to collide with RNA polymerase and cause premature transcription termination (Qi et al. 2013; Bickard et al. 2013).

 

30 Responses to Transcriptional Control

  1. Catie says:

    thx for the research —> Phantastic Phenotypes

  2. Max says:

    Yeah thx petah

  3. EnduroForce says:

    Thankfulness to my father who stated to me about this website, this webpage
    is actually remarkable.

  4. Ziane says:

    Beautiful article, Thank you!

  5. ziane.ziane says:

    I found so many interesting stuff in your blog especially its discussion.

  6. Paulojimmathew says:

    Usually I never comment on sites, but this one is so good. I just want to congratulate your works. Keep it up บาคาร่าออนไลน์

  7. Syntia Silvientos says:

    I have to admit that you have shown a great inspiration in this wonderful site. I understood that we should teach a tactical approach in every learning opportunity that we received even in our own ways of learning before teaching technical skills and showing the result to others to come up for successful and contented result and through this i was able to modify each project that i have specially this one: visit

  8. Paulojimmathew says:

    Much obliged. Thanks! , dude. As usual, you come through like a champ! I like your work. บาคาร่าออนไลน์ มือถือ

  9. Dienneti says:

    Very interesting content!

  10. Andrew White says:

    Hi, I really enjoyed reading your post, and hope to read more. Thank you so much for sharing this.

  11. A basic site you have here… It’s trying to find splendid forming like yours these days.

  12. AMK says:

    Great artcile post,Thank you very so much. INFLUENCER MARKETING AGENCY

  13. العثور بسرعة على اتجاه القبلة في برشلونة. أسهل طريقة لمعرفة اتجاه القبلة في المنزل. استخدم الخرائط على الإنترنت للعثور على القبلة.
    https://www.al-qibla.net/es/barcelona-qibla-finder-1888.html

  14. Hibbah says:

    melhor site de notícias sobre finanças e notícias sobre gamer, jogo free fire, cartão de crédito, empréstimo, etc… harisewell.com

  15. JAKO says:

    Научный портал Fun Sci Club – оплот гибкого критического мышления без предрассудков и мракобесия. Никакой псевдонауки. Только достоверная информация. наука и техника

  16. This article presents clear idea designed for the new people
    of blogging, that actually how to do running a blog.

  17. Mansionbet ไม่ผ่านเอเย่นต์ วอเลท ไม่มีขั้นต่ำ มีครบทกค่ายเกมให้ท่านได้เลือก
    พวกเราเป็น M88 ไม่ผ่านเอเย่น
    วอเลท ไม่มีขั้นต่ำ กำลังเป็นที่ชื่นชอบเพิ่มมากขึ้น เพราะว่าพวกเขาให้สิทธิคณประโยชน์เสริมเดิมแก่ผู้เล่นเยอะมากเมื่อเปรียบเทียบกับเว็บธรรมดา
    เว็บ สล็อดเว็บตรง ไม่ผ่านเอเย่นต์ ซึ่งเป็นอีกกระแสหนั่งในโลกของเกมออนไลน์ สล็อตเว็บตรง กำลังเป็นที่ชื่นชอบมากยิ่งขึ้นสำหรับผู้เล่นที่ถูกใจเล่นเกมรวมทั้งอยากหาเว็บที่ปลอดภัยเพื่อเล่นสล็อตกับ สล็อตเว็บไซต์ตรง เว็บตรงไม่ผ่านเอเย่นต์
    วอเลท ไม่มีขั้นต่ำ

  18. tv8 says:

    Tv8 kanali gibi survivor programini sunan tv kanallari canlitv adresinde. Kesintisiz ve donmayan tv yayinlarini birlikte yasayin

  19. I read this article completely about the difference of most
    up-to-date and previous technologies, it’s amazing article.

  20. APK indir says:

    APK indirmenin kolay adresi

  21. In fact when someone doesn’t be aware of afterward
    its up to other people that they will assist, so here it happens.

  22. Estelle says:

    The brain personal injury lawyer los angeles yelp (Estelle) legal representatives
    at DE CARO & KAPLEN, LLP.

  23. Fornax Software says:

    Günümüz sanayide şuan yaygın olarak kullanılan bir diğer şey ise endüstriyel fırınlar.

Leave a Reply

Your email address will not be published.

Looking for something?

Use the form below to search the site:


Still not finding what you're looking for? Drop a comment on a post or contact us so we can take care of it!

Visit our friends!

A few highly recommended friends...

    Archives

    All entries, chronologically...

    Set your Twitter account name in your settings to use the TwitterBar Section.