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We narrowed to 993 results for: Des

Showing: 181 - 200 of 993 results

  1. CRISPR Plasmids - Plants

    Type
    Collection
    ...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use in ... to a genomic DNA locus and encodes a primer binding site and the desired edits on an RT template. ID ...CRISPR expression plasmids that have been designed for use in plants. CRISPR... can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...gene expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter...

  2. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific... tool for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring ...iGECInano plasmids here!   Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared...intermediate affinities from 40 to 140 mM. These new designs greatly broaden the options for potassium biosensors...further engineering. Figure 5: Structure-guided design and genome mining both led to new sensors with ...

  3. Filming Science Videos in the Age of Social Distancing

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    Blog Post
    ...Good as a WFH desk, and as a video control center.   Cycle 2: Zooming Forward Design: We produced ... So, much like SynBio, it was time to start our Design-Build-Test-Learn cycles. If you’re not familiar...again. Here’s that process!   Figure 1: The Design-Build-Test-Learn cycle adapted for video making...through. I know cameras, but we needed someone else. Design: Jennifer thankfully volunteered, and we decided...here, the operator is in a different room.   Design: The previous cycle made me realize we needed to...restrictions in place, most importantly, masks.  Besides the muffled voices beneath masks, humans rely on...reach for Addgene.  So, onto research! I needed to design a system that would allow me to set up a camera...

  4. Plasmids 101: Common Lab E. coli Strains

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    Blog Post
    ...suggests there are hundreds. This only includes general lab strains designed for subcloning or protein expression...was updated on Nov 14, 2017. You've worked hard designing your plasmid – you carefully selected the optimal...Dr. Theodor Escherich, the scientist who first described them in 1885. E. coli are mainly found in the ...commercial lab strains of E. coli used today are descended from two individual isolates, the K-12 strain ...mutations found in E.coli strains Gene(s) Description Functional Consequence dam DNA adenine methylase... fertility plasmid. A low copy-number plasmid, encodes proteins needed for bacterial conjugation. Genes...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular strains...

  5. Tetracycline Inducible Expression

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    ...promoters (sometimes generally called Ptet) have been designed by combining tet O sequences with elements like...plasmids use "TetO" or "Tet-O" in their name or description, meaning they contain a TRE or tet O and can ...as much flexibility and control as a dual vector design, but it is easier to implement and ensures that...transduction (transient or stable) may depend on the desired sensitivity, level of induction, or amount of background...expression of your gene of interest. ID Plasmid Description Transactivator Promoter PI 21915 Tet-pLKO-puro...Lentiviral Tet-On plasmid for easier cloning and design of inducible shRNA expression, with puromycin selection...transactivator for your tetracycline system. ID Plasmid Description Transactivator PI 26429 pLenti CMV rtTA3 Blast...

  6. Viral Vectors 101: Biosensors

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    Blog Post
    ...have pre-designed plasmids for these systems! Ion sensors One common biosensor class is designed to monitor...all have specific sensors designed for them. For example, this class includes tension monitors associated...interest for many disciplines! Most of the sensors designed for calcium generate a fluorescent output, which... fluorescence in the presence of calcium. This includes FRET-based sensors such as Twitch. While calcium.... Physiological factors Unlike the previously described sensors, this class measures either factors as...to use them, you’ll need to use the vectors (or redesign the tools). It’s a good thing, though! Viral delivery...

  7. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...please improve your plasmid maps!" After thoughtful design, vetting, and tweaking, we’re excited to announce...identifies plasmid features that differ by only a few nucleotides, such as the fluorescent proteins EGFP and mEGFP... software. In fact, SnapGene’s feature library includes nearly 3x as many features as could be detected...greater than our previous mapping software and now includes Type IIS restriction enzymes that are commonly...the gene, the name of its gene product, and a description of its function. You can also click one location...enzymes that cut the given sequence. The table includes enzyme names, the locations where the enzymes ...features detected in the plasmid sequence. The table includes feature names, locations, sizes, colors used to...

  8. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...discussed methods. As described above, you design two primers, each containing the desired mutations, to amplify...make a point mutation, you design two oligonucleotide primers with the desired mutation, then use those ... figure above shows guidelines for primer design; a desired insertion should be present in both primers...vitro recombination. Across all modifications described in Trehan et al., REPLACR-mutagenesis’ median ...

  9. Zinc Finger Consortium: Nuclease Expression Vectors

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    ...This page lists plasmid vectors designed to express nucleases in different cell types. Zinc finger arrays...Expression Vectors This page lists plasmid vectors designed to express nucleases in different cell types. ...zebrafish (can be used as a substitute for pMLM335 described in Foley et al., PLoS One 2009) pMLM292 (for expression...zebrafish (can be used as a substitute for pMLM336 described in Foley et al., PLoS One 2009) pMLM800 (for expression...Methods 2011 ) in mammalian cells or zebrafish as described by Moore et al. PLoS One 2012 and Cade et al.,...

  10. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...coveted plasmid with no substantial delays. Design, design, design Before diving into the experimental work...experimental work. Fragments generation After you’ve designed your cloning scheme, you’re now ready to generate... to use 1-2 ug of the vector for digestion. PCR Design primers that overlap at least 24 bp with the template...from a large genome, you can recheck your primer design or optimize your primers or PCR conditions. Purifying...second and more detailed look at your experimental design. Full lawn of cells. The antibiotic in your plate...complementary to standard sequencing primers. If not, design primers to sequence your insert. When you get your...

  11. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

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    Blog Post
    ... model to help them construct “designer” channelrhodopsins with desired characteristics. The model was...Viviana Gradinaru’s lab used machine learning to design new channelrhodopsins that are more light sensitive...constrained and poorly understood. It’s hard to design new channelrhodopsins with high-throughput proteins...Using machine-learning to overcome channelrhodopsin design challenges To overcome these challenges, the Arnold...experiments. The sequences in this library were designed using structural information about parental channelrhodopsins...existing channelrhodopsins CoChR and/or ChR2(H134R). Designer channelrhodopsins paired with a systemic AAV allow...Find our chemogenetics and optogenetics science guides ...

  12. Important Considerations When Using AAVs

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    Blog Post
    ...utilizing chemogenetics (Designer Receptors Exclusively Activated By Designer Drugs, DREADDs) and optogenetics...beg the question: is axonal transport desirable or not desirable? If diffuse gene expression is tolerated...examining the use of DREADDs (Designer Receptors Exclusively Activated By Designer Drugs) to determine the ...to their Biosafety Level-2 or 2+. Please Note: Despite the fact that AAVs themselves are not pathogenic... CNS tropism (4). If axonal transport is not desirable, transport can be reduced by injecting a smaller...miRNAs), short hairpin RNAs (shRNAs), antisense oligonucleotides (ASOs), zinc finger nucleases, and many others...

  13. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

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    Blog Post
    ...Marraffini’s lab took advantage of this lethality to design the first synthetic bacterial CRISPR system in ...gRNA cassette for plasmid curing through "self-destruction" after induction with L-rhamnose and aTet. CRMAGE...citrea scarless editing plasmids Sheng Yang’s lab describes a two-plasmid system that combines recombineering...Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes Cas9, a tracrRNA, and a CRISPR array, while pCRISPomyces...editing efficiency, perhaps due to its condensed design. For both systems, custom spacers/gRNAs are easily...et al., 2013), the Qi lab employed a gRNA-based design more familiar to those using CRISPR in eukaryotes...plasmid tools that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab...

  14. Viral Vectors 101: Chemogenetics

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    Blog Post
    ...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 ... Gomez J, Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson S (2019) Ultrapotent chemogenetics...Conditional expression and signaling of a specifically designed Gi-coupled receptor in transgenic mice. Nature...

  15. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...that requires modest gene activation. Synergistic Activation Mediator (SAM) Description SAM uses specially...-component system dependent on gRNA design (SAM) or peptide design (SunTag). This streamlines its delivery...genes across diverse cell lines.   dCas9-VP64 Description CRISPR activation can occur through fusing dCas9...requires a relatively simple construct, it exhibits modest levels of gene activation, with some genes experiencing...VPR and SunTag) (Chavez et al., 2016). SunTag Description Rather than using a single copy of VP64 per each...expressed consistently throughout cells. VPR Description VPR fuses a tripartite complex with dCas9 to ...

  16. Synthetic promoter AAVs for cell-type specific expression in retinal cells

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    Blog Post
    ...expression in the retina.  Four approaches for designing synthetic promoters  To build a library of synthetic...synthetic promoters, the Roska lab used four different design strategies: Strategy #1 (ProA): Create promoters...promoters with the help of epigenetic analysis and designed using stretches of DNA that were transcriptionally...mice. Figure 1: The four strategies used to design synthetic promoters. Legend: TSS: transcription...class of cells  Of the four different promoter designs, promoters from strategy #4 had the highest rate...here.  Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular...research for gene therapy. The Roska lab hopes to design improved versions of their current promoters, as...

  17. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

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    Blog Post
    ... 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...

  18. Mammalian RNAi Tools

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    ...target sequence and RISC cleaves and degrades the mRNA. RISC includes argonaute protein 2 (Ago2), which ...RISC). RISC unwinds the double-stranded siRNA and degrades one of the strands. The remaining single-stranded... binding of RISC. For more information on shRNA design and delivery, see resources below. Figure 1: Overview... Trono Lab Lentivectors Toolbox Optimized shRNA design in pSico with pSicoligomaker (Andrea Ventura lab...Taxman, D. J. (2010). Short hairpin RNA (shRNA): design, delivery, and assessment of gene knockdown . Methods...

  19. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...of screenshots to highlight changes to both the design and functionality of searches. A better way to ...posts, protocols, and more. Selecting crispr in Guides will return links including our CRISPR guide.  ... isn’t new to Addgene’s plasmid search. The new design of the plasmid search page moves the filters from...Lentiviral’ which eliminates most of the plasmids designed for other uses. I could look through these resulting... information about the subject, link to related guides and protocols, and provide easy to sort tables ...to the CRISPR Resources page: The page links to guides, our CRISPR eBook, plasmid kits, and curated pages...by a convenient table of plasmids specifically designed for this purpose: Still stumped? Ask an Addgenie...

  20. CRISPR 101: Making a Knock-In Cell Line

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    Blog Post
    ...harnessing the homology-directed repair pathway, designing the best donor DNA, and avoiding common mishaps...knock-in. Then choose which Cas enzyme to use and design a gRNA to where you would like to introduce the...sequence to introduce, which is done through the design and use of a donor DNA molecule. Your donor molecule...strand break by homologous recombination.    Designing donors for HDR The basis of the HDR pathway relies...used as well. Below are some considerations for designing donor DNA for successful HDR events. CRISPR cut...need to be used due to synthesis limits of oligonucleotides. These donors traditionally have larger homology...or part of the guide’s recognition sequence is destroyed via mutation. By introducing a mutation in either...
Showing: 181 - 200 of 993 results