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

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We narrowed to 1,045 results for: ins;

Showing: 321 - 330 of 1045 results

  1. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...upstream of either NGA or NGC PAM sequences (Kleinstiver et al. 2015). From these screens, they identified...mutations found in the REC2, REC3, and PAM interacting domains and allows for expanded PAM recognition as well...off-target activity (Hu et al., 2018).  In 2020, the Kleinstiver lab reported the development of near-PAMless ...the CRISPR toolbox The isolation of novel CRISPR proteins has and will continue to dramatically increase...which makes it easier to target AT-rich genomes (Kleinstiver et al., 2016). Type VI CRISPR systems, which ...Wolfe SA, Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome... plasmids from this publication at Addgene. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng...

  2. Hot Plasmids - August 2020

    Type
    Blog Post
    ...cleave affinity tags from purified recombinant proteins at small and large scale.  This new technique ...technique involves selection in the cytosol (instead of on the yeast extracellular surface) which enables tracking...demonstrated to work for evolution of other target proteins and might be useful for your studies.  Find the... Angela Abitua Genetically encoded fluorescent proteins are often used to label  specific structures, ...within cells. However, sometimes these fluorescent proteins might not localize to the right places, which ...to improve nuclear localization of fluorescent proteins, they found that adding a triple nuclear localization...in the cytosol of E. coli, however the nanobody proteins tend to get stuck in inclusion bodies. Takayuki...

  3. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...Strong insect promoter from baculovirus Constitutive  Commonly used in expression systems for insect cells...to replicate plasmids and make sure the cells maintains it. Here, we'll cover the promoter, which is necessary...responsible for initiating the transcription of your insert into RNA. In practice, the term "promoter" describes...regulatory factors: enhancers, boundary elements, insulators, and silencers; however, some "leakiness" of ...Strong hybrid mammalian promoter Constitutive  Contains CMV enhancer, chicken beta actin promoter, and...with Tetracyline or its derivatives.  Typically contains a minimal promoter with low basal activity and... promoter. Ac5 General expression mRNA Strong insect promoter from Drosophila Actin 5c gene Constitutive...

  4. Spurred by COVID-19, Addgene Bridges Plasmid Sharing Between Academia and Industry

    Type
    Blog Post
    ...                           California Institute of Technology             Johns Hopkins University                           ...                           National Institute for Physiological Sciences Broad Institute                                       ...for tagged and untagged expression of SARS-CoV-2 proteins in bacteria, yeast, and mammalian cells. We teamed...from the Michael J. Fox Foundation are used in Parkinson’s research.  The extensive resource from the Structural... determine the three dimensional structures of proteins.  The RESOLUTE Consortium collection enables research... collection, a ‘One-Stop Shop’ for Fluorescent Proteins, and plasmids from several labs at Florida State...

  5. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...problems? Instead of directly fusing a fluorescent protein to your protein of interest, you instead fuse it...possibility of long-term imaging studies. If you instead overexpress your protein at higher levels, you ...is at a very high concentration. Overexpressed proteins also have the potential to form aggregates and...to the synthetic SunTag scaffold. This scaffold contains 10-24 copies of the short epitope GCN4. GCN4 recruits...SunTag, they attempted single-molecule imaging deep inside the cell, in the nucleus and cytoplasm. Again, ... of SunTag compared to traditional fluorescent proteins Improved brightness and signal-to-noise ratio... fluorescent proteins...

  6. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ...alarming results are due, at least in part, to insufficient and non-standard record keeping despite scientist...to capture ideas, write down certain notes or brainstorm, but we have to admit that paper-based record...functionalities that are hard to imagine on paper, such as instant search through all saved data and automated writing...maintenance and support.” Besides that, academic institutions tend to think about the long term, so they need... once they establish the system within their institution. In the study, respondents were also asked about...providers used to supplement their paper notebooks. Instead, all of the important information is consolidated... In addition, if you are working for a large institution or company, site licenses might be available ...

  7. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    ...own machinery to replicate the viral genome and proteins. This is, for the intrepid researcher looking ...Virus structure While all viruses have a genome inside a protein shell known as a capsid, some also have...can either follow central dogma (gDNA to RNA to proteins) or require an intermediary DNA replication step...transcribed into mRNA, directly translated into proteins, or, in the case of retroviruses, reverse transcribe...transcribed into mRNA before being translated into proteins. RNA viruses replicate their genomes using a variety...egg sac. (Okay, some enveloped viruses bud out, instead of lysing, but they do generally still kill the...replicative ability. The remaining particle, which retains the protein coat (and envelope), has a “gutted”...

  8. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    ...please use one of these methods to ensure your insert retains the 3' A overhang: Use a mixture of proofreading...below, the “A” overhang on the blue PCR product insert comes from using Taq polymerase for the amplification...these kits also often have the topoisomerase site inserted into a beta-galactosidase cassette allowing a ...clones. Once you introduce your 3’-end “A” overhang insert, the magic of TOPO cloning happens. Basic procedure... Colonies: You can confirm the presence of your insert by PCR, restriction digest, or sequencing. Pro ...concentration or you are cloning an extremely large insert. Since the standard ligation reaction is fairly...PCR Use REPLACR Mutatgenesis to Easily Generate Insertion and Deletions in a Plasmids Learn How to Verify...

  9. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    ...Plasmids 101: E. coli Strains for Protein Production Addgenie Julian Taylor-Parker explains why you might want...Homology Directed Repair Addgenie Chari Cortez explains some of the basic science behind homology directed...production and gives a quick rundown of some of the strains you can use to express your protein of interest...find a successful clone? Addgenie Jessica Welch explains how Blue-White screening could be a potential ...Addgene Science Communications Intern Mary Gearing explains how Cas9 homolog, Cpf1, may make it easier to ... able to help you. Head Addgenie Joanne Kamens explains what career coaches do and offers up some career... to use with CRISPR as the cargo. Mary Gearing explains some of the clever ways to successfully combine...

  10. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...Oliver Griesbeck of the Max Planck Institute for Neurobiology has been working on genetically encoded...used for ratiometric in vivo imaging. Griesbeck explains, while these Twitch sensors are always bright,...particular advantage for observing events taking place inside cells that are on the move, such as T lymphocytes...Eindhoven has similar interest in FRET-based sensor proteins, particularly for the intracellular imaging of... should require the simple swapping of binding domains, in practice optimizing the sensors involves plenty...solution: they devised self-associating fluorescent domains whose association is disrupted in the presence ... of mOrange2 and mCherry as donor and acceptor domains. He says, these new colors might now enable observation...
Showing: 321 - 330 of 1045 results