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We narrowed to 79 results for: tet off

Showing: 41 - 60 of 79 results

  1. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...) the consequences to your experiment if there’s off-target expression.   Once you know that, you’re ready...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig. 3) - for ... two astrocyte selective promoters demonstrating off-target expression in both oligodendrocytes (Olig2...promoter allows you to decide when to turn on, or turn off, gene expression by administering an inducing agent...  Finally, the risks of potential toxicity and/or off-target effects (e.g. recombinase-dependent vectors...Ubiquitous promoters  These often-strong promoters offer the most flexibility for expression in a large number...2002). Lack of an Immune Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term...

  3. What's New in CRISPR - May 2019

    Type
    Blog Post
    ... in the rat APOBEC1 that decreases the number of off-target RNA edits by >390 fold and >3,800 fold in .... coli genome-wide CRISPR inhibition Covalent tethering of DNA template to Cas9 SECURE base editors Nme2Cas9...genome-wide CRISPR inhibition library Covalent tethering of DNA repair template to Cas9 Homology-directed...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... temporally control which genes are turned on or off in a given area and the can do so in a very specific...gene expression studies, which are evinced by the Tet system and visualization of single molecules via ... which allow you to easily and reversibly switch off and on your favourite signalling pathways (Figure...of the ribosome during translation, which knocks off all mRNA binding proteins as it travels along the...for Translation RNA Imaging by Coat protein Knock-off, is made up of three components: a GFP protein that...NLS-MS2-RFP) the reporter mRNA, which contains the tethering sites for the fluorescent proteins and an inducible...however, remain bound to the mRNA since they are tethered AFTER the stop codon. Thus a non-translated mRNA...

  5. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...some challenges still exist. Nucleases may cause off-target effects on regions of the genome with similar...either induce or repress Cre expression using TetON or TetOFF systems. So now hopefully creating a genetically... the cell can occur. Similar systems include tetracycline or doxycycline mediated activation of Cre to...

  6. Let There Be LITE Plasmids

    Type
    Blog Post
    ...designed to bind specific genes and turn them on or off, literally at the flip of a (blue) light switch. ...chromophores flavin-adenine dinucleotide and methyltetrahydrofolate, which are universally abundant. Second...apply LITEs for the transformation of neurons, she offers a tip: save yourself some time by making an AAV1...

  7. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ... GFP-binding nanobodies, the project really took off. Unlike most antibodies, nanobodies, which occur ...DNA-binding domains, including the commonly used LexA and rTetR systems such that they activate gene expression ...advantages. T-DOGGs can be made drug-inducible using the rTetR system, but Cre-DOG lacks the toxicity that can ...

  8. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... Wheeldon FgH1tUTG 70183 Mammalian/Lentiviral H1-Tet none S. pyogenes Herold lentiGuide-Crimson 70683 ...Zhang pRS416-dCas9-Mxi1 + TetR + pRPR1(TetO)-NotI-gRNA 73796 Yeast pRPR1(TetO) yes, interfere S. pyogenes... 65006 Bacteria BsaI yes, interfere S. pyogenes Koffas BPK764 65767 Bacteria BsaI yes, cut S. pyogenes...yes, nick S. pyogenes Duchek pAC5-dual-dCas9VP48-sgTetO 48237 Mammalian BbsI yes, activate S. pyogenes ...Joung AAV:ITR-U6-sgRNA (Backbone) PCB-FlPO-WPRE-syntetisk pA-UTR 68347 Mammalian/AAV none S. pyogenes Elverlov-Jakobsen...62315 Yeast none S. pyogenes URA3 Lim, Qi pRPR1(1xTetO)_gRNA_handle_RPR1t 62966 Yeast HindIII none S. ... Mammalian/Lentiviral hU6 none S. pyogenes Puro Moffat lenti sgRNA(MS2)_puro backbone 73795 Mammalian/...

  9. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ... on-target activity to siRNA, but with far fewer off-target effects! This characteristic probably explains... by DNMT3A or MQ1, and cytosine demethylation by Tet1. RNA Targeting: This page contains Cas13 constructs...

  11. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...detection methods lack spatiotemporal control or an “on/off switch.” This makes them great tools for providing...speed of labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein...Split-BioID, Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting...

  12. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ... underwent affinity screening to select for slow off-rate binders. The best candidates were validated ...where gRNA expression is induced by anhydrous tetracycline. Three new anti-CRISPRs were identified that...Janelia Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes ...-mRuby from Liqun Luo’s lab expresses membrane tethered GFP to enhance axonal labeling and synaptophysin-tagged...

  15. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...another, or their fluorescence can be switched on and off reversibly. This ability to modulate fluorescence...PA-FPs come in 2 flavours- monomeric and tetrameric. Tetrameric PA-FPS are better for whole cell imaging...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...UV-Violet Cyan/ Green >2,000 Medium 490/ 511 Kaede Tetramer UV-Violet Green/ Red ~2,000 High 572/ 580 tdEos...UV-Violet Green/ Red N/A High 573/ 584 KikGr Tetramer UV-Violet Green/ Red >2,000 High 583/ 593 mKikGR...Monomer Blue Dark/ Red 20 Low 572/ 610 FP595 Tetramer Green Dark/ Red 70 - 1000 Medium 590/ 600 ...non-invasive labelling, but in contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity...

  16. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...generate a knockout or edit. This can help reduce off-target activity. Deleting a large region of the genome...by L-arabinose and anhydrotetracycline (aTet), respectively. pMAZ-SK contains an aTet-inducible gRNA and...coli multiplexing plasmid is available from the Koffas lab. Scaling up your multiplexed gRNAs with Golden... on a separate plasmid. Multiplexing in E. coli Koffas Lab CRISPathBrick Multiplex Plasmid: This system...self-destruction" after induction with L-rhamnose and aTet. CRMAGE is much more efficient than traditional ...plasmids from this paper at Addgene. 3. Sakuma, Tetsushi, et al. “Multiplex genome engineering in human...

  17. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...binding partners does it have and how do they come off and on in a cellular environment? If these are pressing... nonfluorescent until they bind a 6-residue tetracysteine tag (sequence: CCPGCC) that can be inserted ...interest, resulting in a C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small...et al. "Hairpin structure of a biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal...

  18. CRISPR Guide

    Type
    Guide
    ...increase specificity, and decrease off-target effects Sniper-Cas9 - less off-target activity; compatible with...cleaving only one strand of target dsDNA. Off-target effects or off-target activity Cas9 cleavage at undesired...of partial homology throughout the genome, called off-targets, that can impact your experiment. There are...DSB within the target DNA, it’s unlikely that two off-target nicks will be generated close enough to cause... mutating specific amino acid residues to reduce off-target editing. Some mutations disrupt interactions...method, increased fidelity enzymes generate less off-target editing than wild type Cas9. Examples of increased...proofreading and discrimination evoCas9 - decrease off-target effects xCas9 3.7 - mutations in multiple ...

  19. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...functional knock-out via frameshift and minimize off-target effects. The library was cloned into the Liu...inserting OsTIR1 into the "safe harbor" AAVS1 locus (a tet-inducible OsTIR1 plasmid is also available). Second...actin 5C promoter. Addgene’s fly library web page offers more details and protocols, and the Liu lab’s OXfCRISPR...unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many of which ...context-dependent human fitness genes, the laboratory of Jason Moffat at the University of Toronto has created the Toronto...demonstrate the effectiveness of their library design, Moffat laboratory members conducted multiple screens in...hits. This work has been published in Cell. The Moffat laboratory maintains a website for the TKO library...

  20. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology...CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. It can be very time consuming to ...CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. He has a particular interest in using...
Showing: 41 - 60 of 79 results