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We narrowed to 634 results for: des.2

Showing: 521 - 540 of 634 results

  1. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ... the Cas machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA... reference?  The CRISPR guides used in the screens they analyzed were designed to have a perfect match...African descent have approximately 20% more single nucleotide polymorphisms (SNPs) that map to guides in the...accounted for in the CRISPR guides they used, leading to mismatches between the guides and the lines’ genomes...modify the CRISpick algorithm to prevent guides from being designed that target regions with high genomic...coming through an unexpected source: the CRISPR guides, short pieces of RNA that bind to a cell’s DNA,...x-axis) which SNPs map to thetargeting sequences of guides across 8 CRISPR libraries. Samples are divided ...

  2. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    ...of these issues are not particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges... Zimmer provides several. In Part Three of the book, called simply “Old Science,” he describes the now...teaching and communicating science for over two decades, attempts to answer it in five paragraphs, with...recent resurgence of “Do-It-Yourself Science” described in Chapter 3, and of course the rapidly changing... science.”  The three chapters in this section describe the challenges of trusting science in a “fake ...changed significantly over the last decade. He describes the detection of gravitational waves--a story ...collaboration from scientists around the world. The paper describing the waves’ detection had over 1,000 authors. ...

  3. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...Reports 9: https://doi.org/10.1038/s41598-019-41088-2 Magnus C, Lee P, Bonaventura J, Zemla R, Gomez J, ...DREADDs DREADDs DREADDS, or designer receptors exclusively activated by designer drugs, are G protein-coupled...Relationship Studies for Designer Receptors Exclusively Activated by Designer Drugs. ACS Chemical Neuroscience...Chemogenetics has become a popular tool in the field as it provides a noninvasive method for manipulating cell activity...Roth's and Takafumi Minamimoto's lab called deschloroclozapine (DCZ) shows promise as a CNO alternative ..., Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson S (2019) Ultrapotent chemogenetics for research...Conditional expression and signaling of a specifically designed Gi-coupled receptor in transgenic mice. Nature...

  4. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...integrates into the trnS-leu2 locus.   Figure 2: Introducing Mobile-CRISPRi into bacteria. Top: A ... human gut flora. Guides targeted essential and non-essential genes with two guides per gene. The library...expressing guides that target essential genes were less represented while bacteria containing guides targeting... Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined suite of tools for using CRISPRi...existing bacterial CRISPRi methods. First, its modular design allows for customization with strain-specific promoters...s take a look at the essential gene folA which encodes the target for the antibiotic trimethoprim. When... modular nature allows for the customization of guides, promoters, selection markers, and different versions...

  5. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...split at 175 Arabidopsis homolog of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin...Have no fear, Addgene is here to help!  General Design Figure 1: Two proteins are fused to the ...from the associated paper:   Color Base FP Description Article (Link to Plasmids) PI Blue EBFP2...contains both fragments of split mVenus (aa 155), includes I152L in N-terminal fragment An enhanced recombinant...Dan Mulvihill mVenus Split mVenus (aa 155), includes I152L in N-terminal fragment Constant rate of ...reversible complementation. TagBiFC (split HaloTag) provides maximum flexibility in color labeling with exogenous...single-molecule tracking in live cells.  Controls  When designing experiments, it’s important to include proper ...

  6. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...) 12 (>100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent...insertion site, the corresponding gRNA might be undesirable due to potential off-target sites or low predicted..., resulting in lower overall genome frequency. Despite the progress we’ve made, there is still a need ...bacterial polymerase subunit ω. This construct is designed to activate transcription from the accessory plasmid... stringency and promoting survival of only the broadest-targeting variants. For the final round of evolution...spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing NG, NNG, GAT, and CAA PAMs...scaffold. With an NGG PAM, xCas9 3.7-VPR constructs modestly outperformed SpCas9-VPR in activating transcription...

  7. Rabies and Neuronal Tracing

    Type
    Blog Post
    ... Dec; 18(6): 617–623. Pubmed PMID 19349161.  2. Deshpande A, Bergami M, Ghanem A, Conzelmann KK, Lepier...widely used in neuroscience. In this post, I’ll describe how rabies virus (RABV) can be used in the brain... by modifying the wild-type RABV genome, which encodes a single envelope protein called rabies glycoprotein... neurons integrated into the brain over time (Deshpande et al., 2013). The study found that at early time...Pubmed PMID 26190977. 6. Osakada F, Callaway EM. Design and generation of recombinant rabies virus vectors...

  8. Of Myc and Men

    Type
    Blog Post
    ...promising monoclonal antibody with high specificity. (2) Narrow down the specific epitope that your antibody...append their proteins of interest with unrelated peptides that already had high quality antibodies available...existence of high quality antibodies for well defined peptides. Munro and Pelham’s first published peptide tagging... tumorigenesis.  Using two different synthetic peptides corresponding to linear sequences within the middle...that perfect. One of the known limitations was described by Schüchner, et al., who published a paper showing...works and allow users to better troubleshoot and design experiments. It turns out that even with the most...

  9. Hot Plasmids February 2024

    Type
    Blog Post
    ...overview of these new variants:   Figure 2: Inner mitochondrial membranes imaged with structured... They fused the Cas enzyme to cell-penetrating peptides and co-incubate it with an “assist peptide” that...with a 30-minute incubation, even with multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused...all applications. Addgene’s plasmid collection includes over 10,000 plasmids with a FLAG tag (!), but ...recombinant antibodies as well? Our collection includes a mouse and a chimeric rabbit (original mouse ...

  10. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...PMID: 25538079. PubMed Central PMCID: PMC4353587. 2. Stephen, Andrew G., et al. "Dragging ras back in ...die from cancer-related complications each year. Despite these grim statistics, new strategies for attacking...cancers, especially those of the pancreas and colon. Despite the prevalence of Ras mutations in a variety of... enabled researchers to begin discovering and designing molecules that are showing some promise in their...in the screenshot below). Currently, the page includes interactive maps for the Ras, mTor, and p53 pathways...

  11. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...PMID: 27151215. PubMed Central PMCID: PMC4938170. 2. Zuris, John A., et al. "Cationic lipid-mediated delivery...transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains a site for inserting a custom...targeted approach using RNPs has recently been described in vitro, with the hope of eventually using this...expertise. Final thoughts The Cas9-gRNA RNP method provides scientists with an additional means to deliver...other CRISPR delivery techniques Find tips on gRNA design Resources on Addgene.org Browse all CRISPR plasmids...

  12. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ... replication competent virus [usually HIV-1]; and 2) the potential for oncogenesis through insertional...those seen when using a plasmid backbone that was designed specifically for transient expression (find some...expression of viral constructs is a useful tool as it provides a means to quickly check that a construct is functional...3rd-generation transfer plasmids have a chimeric 5'LTR that includes a CMV or RSV promoter as well as a portion of ...those three genes plus tat. The envelope plasmid provides a heterologous envelope for pseudotyping and is...

  15. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...You’ve probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering...non-coding RNAs (lncRNA). lncRNAs are over 200 nucleotides in length and lack an open reading frame, so ...transcription machinery of a target gene or DNA that codes for ncRNA. CRISPRa, on the other hand, recruits ...which targets RNA instead of DNA and selectively degrades RNA molecules. This system is often used to knock...

  17. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ... Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed...resistant to disease. They also devise biosensors designed to elucidate the molecular interactions that go...things for day-to-day life in a plant lab, so it includes everyone’s favorite promoters, plus a few new ...

  18. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis...adapted "self-cleaving" 2A peptides into their muliticistronic vectors. These peptides, first discovered in ...plasmids) are designed to have one or more of the genes replaced.  Additionally, 2A peptides could be PCR-cloned...PDF that you can save on your desktop. The Plasmids 101 eBook is designed to educate all levels of scientists...incorporated into plasmids for research purposes are described below. IRES Elements Translation in eukaryotes...the specific genes cloned into the vector.3  2A Peptides To overcome some of the disadvantages of the IRES...self-cleaving" is not entirely accurate, as these peptides are thought to function by making the ribosome...

  19. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...overhang twice; (2) avoid palindromes; (3) no overhangs with the same three nucleotides in a row; (4) no...rules of thumb for designing your overhang sets. But are those rules the best way to design GGA overhang sets...data-optimized assembly design (DAD) to build out three tools to help researchers design high-fidelity Golden... no more than two nucleotides in the same position; (5) avoid overhangs with either 0% or 100% GC.) In...the traditional rules of thumb for overhang set design allowed for high-fidelity assembly for sets of ... overhangs that violate some of the traditional design rules) for successful high-complexity Golden Gate...with a high potential for mismatches, so you can redesign individual pieces as needed to improve fidelity...

  20. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...Central PMCID: MC4262738. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and ...These libraries utilize optimized sgRNAs that were designed based on the rules identified in Doench et al....1,000 non-coding control sgRNAs. This library was designed to improve on-target activity while reducing off-target...with 6 sgRNAS, highlighting the benefit of sgRNA design in CRISPR libraries. Thus, the Brunello library... that if an essential gene plays a role in the desired screening phenotype, it may be missed because it's... the sgRNA metrics, confirming again that sgRNA design plays a significant role in library efficiency....CRISPRa, sgRNAs were targeted between 150-75 nucleotides upstream of the TSS. Similar to the Brunello ...
Showing: 521 - 540 of 634 results