Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 1,023 results for: nts

Showing: 41 - 60 of 1023 results

  1. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...Sequence NGG CRAA (R = A or G) Spacer Length 20 nt 22 nt Temperature Range for Optimal Activity 35 - ... organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs, some bacteria; but not...s love of heat in two ways. The first set of experiments tested the ability of GeoCas9 RNPs to be incubated... following heat treatment. The second set of experiments tested the ability of GeoCas9 RNPs to cleave ... we haven’t thought of yet? Let us know in the comments below. And if you want to give GeoCas9 a try, ...

  2. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ... by the Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant...68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD plasmid...identifies a staggering ~250,000 phosphorylation events in the proteomes of higher mammals (2). How can... the importance of any of these phosphorylation events on the activity of a given protein? Classical methods...5). Optimization of this system defined three components required for translation (Figure 1): A tRNA ...the pSer-incorporated polypeptide. These three components were shown to effectively incorporate phosphoserine...inactive without requisite dual phosphorylation events on its activation loop, but can be made constitutively...

  3. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...efficiencies of multiplex editing at each site. A 75-nt human cysteine tRNA stood out from their initial ...array, DAP arrays use tRNA itself, as short as 75-nt, to both drive and process the tRNA-gRNA arrays, ...both efficiency and multiplexity, with these constraints? The Gao lab first thought to enable MBE by ...the DAP array, the Gao Lab packaged MBE and MPE elements for deliveries via adeno-associated virus (AAV...

  5. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...CRISPR/Cas system makes use of 3 components, a guide RNA (gRNA) of about 125 nt that specifies the target, ...the mouse genome is made up of 3 x 109 nucleotides (nt), and encodes 23,000 or so genes. It would be great...the gRNA will seek out its target among the 3 X 109 nt of genetic content in the mouse genome and the Cas9...gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together and microinjected into either...eggs. Or, to avoid handling embryos ex vivo, the components can be electroporated into the oviduct of pregnant...CRISPR is so efficient that you can inject the reagents directly into fertilized mouse eggs, circumventing...friends and colleagues while working with these reagents. Other than that, be nice to your microinjectionist...

  6. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...plasmid use 30 nt oligos for cloning but PX330 only uses 20 nt? A16: The reason for the 30 nt region on the...In the pre-processing form, the crRNA contains 30 nt of the target spacer, but then when expressed in ...double nickase for my CRISPR genome engineering experiments? A1: When assessing which nickase type to use...use for your CRISPR genome engineering experiments, consider that wildtype Cas9 with optimized chimeric...efficiency is very high. Q6: How should I design experiments for multiplexed targeting with Zhang lab plasmids...

  7. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ... kb ~3.9kb crRNA/gRNA length gRNA: ~100 nt crNA: ~42 nt Requires tracrRNA? Yes No dsDNA cleavage...Cas9 and Cpf1 CRISPR Nucleases   Cas9 Cpf1 Components crRNAtracrRNACas9 crRNACpf1 Endonuclease...AAV transduction.  In most of Zetsche et al’s experiments, a single vector co-expressing Cpf1 and a crRNA...culture of mouse neurons and in vivo. For these experiments, cells were infected with a 1:1 ratio of two ...editing with CRISPR-Cpf1 is one of the latest developments in CRISPR technologies.  It’s a simple and effective...thoughts about Cpf1 multiplexing, leave them in the comments below.        References 1. Zetsche et al. "Multiplex...

  8. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...)  Editing window ~2 nt  85175  pBK-EE-BE3  SpCas9 (NGG)  Editing window ~2 nt  85176  pBK-YE2-BE3...BE3  SpCas9 (NGG)  Editing window ~2 nt  85177  pBK-YEE-BE3  SpCas9 (NGG)  Lower on-target activity than... than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity...Campbell’s lab added two new mCherry variants to the repository. These variants can be used as tools to learn... Podcast Segment The impact of KLF4 N-terminal variants on iPSC generation Deriving iPSCs (induced pluripotent...transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered ... developed a new AAV approach using regulatory elements that restrict expression to this cell type. To...

  9. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ... made up of microtubules, actin filaments, and intermediate filaments. These structures give the cell ...GFP-cortactin Actin Filaments Cortactin EGFP Anna Huttenlocher 32907 pEGFP-rNFM Intermediate Filaments (neuronal...Intermediate Filaments (neuronal) Nefm EGFP Anthony Brown 11908 pEGFP-N1 alpha-actinin 1 Actin Filaments alpha-actinin...Actin Filaments Cortactin mCherry Christien Merrifield 36201 pLifeAct-mTurquoise2 Actin Filaments LifeAct...pLifeAct_mScarlet_N1 Actin Filaments LifeAct mScarlet Dorus Gadella 85055 pLifeAct_mScarlet-H_N1 Actin Filaments LifeAct ... Actin Filaments ITPKA mTurquoise2 Dorus Gadella 58473 pEGFP-C1 F-tractin-EGFP Actin Filaments F-tractin...mCherry-Lifeact-7 Actin Filaments LifeAct mCherry* Michael Davidson 54610 mEGFP-Lifeact-7 Actin Filaments LifeAct EGFP...

  10. Antibody Guide

    Type
    Collection
    ...antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments - Fab fragments are created... called the antigen binding fragments (Fab). The tips of the Fab fragments contain highly variable regions...Affinity Reagents There are several alternatives to antibodies, ranging from antibody fragments (shown ...Structure Isotypes Antigen Specificity Other Affinity Reagents Handling Visualization Signal Amplification Application...Multiplex Assays Selection Validation Affinity reagents are molecular tools used in the lab to specifically...foreign bodies, are the most commonly used affinity reagents. Antibodies are Y-shaped proteins, naturally produced...interest is introduced to the animal, which then mounts an immune response to it. Once the immune response...

  11. Plant Plasmids and Resources

    Type
    Collection
    ...in monocot and dicot plants - Yao-Guang Liu. Plasmids for prime editing in plants (including rice and ...Addgene's plant resources: Find plasmids for use in plants. Plasmid...tools that were specifically designed for use in plants. These resources include tools for research using...useful toolkits and plasmid collections for use in plants. The tables below highlight plasmids spanning a...selection of plasmids expressing different functional variants of Cas9 and empty gRNA backbones. A few examples...- Yinong Yang. The MoClo CRISPR/Cas Toolkit for Plants includes CRISPR/Cas nucleases, base editors, gRNA...assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner. CRISPR-Cas9 vector with GRF4-GIF1...

  12. CRISPR Guide

    Type
    Collection
    ...multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements, and other Cas...scale edits by integrating segments of DNA flanked by specific repeat elements into the genome. CRISPR-associated...mechanism Engineered CRISPR systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated...mechanisms of CRISPR and some of the ways the CRISPR components are leveraged to achieve specific goals, such...the Cas enzyme. Basic Mechanisms of CRISPR The requirements for a CRISPR knockout assay are simple. You’...Cut) Multiplex Genome Engineering Many CRISPR experiments involve editing more than one gene. In these ...RuvC and HNH, can be altered to reduce cleavage events to a single cut (nick) or no cuts. Cas9 nickase...

  13. Plan Your Experiment

    Type
    Collection
    ...proteins available, and researchers can adapt these experiments for different organisms. This page provides a...CRISPR in your research and setting up CRISPR experiments. We will focus on making single edits using CRISPR...using CRISPR in other organisms. As with most experiments, you first need to have a biological question...you are ready to start designing the specific components (Cas enzyme and guide RNA) for your experiment...genetic manipulations require different CRISPR components. Selecting the genetic manipulation you plan ...plan to perform will help you narrow down which components you’ll need for a given experiment. In the table... window from the PAM sequence, and base editor variants often have different optimal editing windows. ...

  14. Antibody Plasmid Collection

    Type
    Collection
    ...antibody components into plasmids and expressing these in bacteria, mammalian cells, yeast, plants, or insect...size. These recombinant antibody fragments can be used in experiments such as immunoprecipitation and ...antibodies as Fab' fragments, isotype-switched chimeric antibodies, and Fc-silent mutants. Functional diversification...microscopy. Two commonly used plasmid-based antibody fragments are scFvs and single-domain antibodies (also known..., scientists often use plasmids encoding scFv fragments that bind to the GCN4 peptide, from the popular...the generation of chicken monoclonal antibody fragments by phage display. J Immunol Methods. 2000 Aug ...dimerisation and detection of recombinant antibody fragments in the form of scFvs A simple vector system to...

  15. Luciferase Plasmid Collection

    Type
    Collection
    ...luciferase a popular tool for scientists. Regulatory elements such as promoters, enhancers and untranslated ...secreted and more stable, enabling time course experiments, but is too dim for some applications. NanoLuc...Creating ORF-Firefly luciferase gene fusions in plants. Renilla luciferase under the control of a CMV ...white screening can be used for screening of transformants in cloning process. Lynne Postovit 64034 pGL4Luc-RLuc...fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174051 Alberto ...fragment for split-luciferase complementation assay in plants using Gateway cloning. Use with 174050 Alberto ...Transcriptional unit for firefly luciferase expression in plants (GoldenBraid 2.0). Diego Orzaez 68215 pEGB 35S:...

  16. Bacterial Expression Systems

    Type
    Collection
    ...bimolecular fluorescence complementation experiments in plants. The Zinc Finger Consortium Modular Assembly...plasmids that can help researchers plan their experiments using bacterial expression systems. Read below...proteins of interest to transcriptional regulatory elements that will turn on gene expression upon protein–protein...generation of FRET biosensors for ratiometric measurements. The MoBiFc Toolkit (Addgene #1000000222) is...Plasmids Reporter plasmids can be used to detect events and molecules in and around a bacterium. Plasmids...Chromoproteins: Colorful Proteins For Molecular Biology Experiments Return to top Genome Engineering Modifying the...cellular resources otherwise spent on producing high amounts of protein. Whether you're investigating basic ...

  17. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ...C-terminal fragments and placed under the control of different promoters. Expression of both fragments in the... gene expression, and find plasmids for your experiments. Plasmid...Site Canonical Sequence Structure (Length) Common Variants Cre loxP ATAACTTCGTATAgcatacatTATACGAAGTTAT 13...system allows scientists to utilize recombination elements such as Cre to turn off the expression of one ...combined in multiple, independent recombination events in a single experimental system, enabling multi-layered...lineage-specific drivers can also help reduce off-target events and the systemic effects of Cre toxicity. Leakiness...iCre variant, reduces the number of recombination events but can improve specificity in targeted neuronal...

  18. Lentivirus Plasmids

    Type
    Collection
    ... Guide for more information about: Lentiviral components 2nd versus 3rd generation lentiviral systems ...feature of lentivirus used in research is that the components necessary to produce an infectious viral particle...called packaging and envelope plasmids) encode components of the viral capsid and envelope and are used...transfer plasmids along with additional packaging components Trono 63586 psPAX2-D64V 2nd Packaging For making...that encodes genetic material that the researcher wants delivered to specific target cells. This genome ... See article for more cre-lox shRNA expression variants. Jacks 19319 pLJM1-EGFP 3rd CMV-driven EGFP fusion... 11619 pLB 3rd Modification of pLL3.7; Genetic elements known to prevent epigenetic silencing were added...

  19. Tetracycline Inducible Expression

    Type
    Collection
    ...tetracycline-regulated gene expression and find plasmids for your experiments using tet- or dox-inducible systems....system has been engineered with a variety of improvements. A synthetic Tet Response Element (TRE) typically...been designed by combining tet O sequences with elements like the H1 RNA promoter. And, most importantly...Tet-Off and Tet-On. Read on to learn more about the components of tet systems and how to use them, or skip ahead...activation domain). For simplicity, other promoter elements are not shown. Tetracycline Off (Tet-Off) The ... Systems Subsequent efforts have improved the components of these systems for better efficiency, sensitivity...). rtTA3 and Tet-On 3G : further improved rtTA variants with lower background activity and greater sensitivity...

  20. Distribution to Industry

    Type
    Collection
    ...to be diverse in topology, size, and isoelectric points. Pooled Libraries Pooled Library name Type PI Description...payment, and billing. Industry Material Transfer Agreements Most Addgene orders must be accompanied by a ...protection for material providers. Addgene's deposit agreements allow Addgene to distribute plasmids on behalf...behalf of the depositing institutions. These agreements indicate the language which we are allowed to use...use for distribution. Please see the Sample Agreements on our Technology Transfer Information page for ...authorized signatory is someone who can execute legal documents on behalf of the recipient institution. Once the...addgene.org . After we review and approve the MTA documents, your order will be processed and shipped. Industry...
Showing: 41 - 60 of 1023 results