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We narrowed to 770 results for: VARS

Showing: 161 - 180 of 770 results

  1. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...that don’t utilize NGG   There are plenty of Cas9 variants which utilize alternative PAMs. These alternative...rounds of engineering, they developed the SpRY Cas9 variant (Walton, et. al, 2020). SpRY uses NRN PAMs and ...efficiency and less reliability. Notably, the SpRY variant didn’t display higher off-target effects than SpCas9...experiments. Between SpRY and several of the other Cas variants described, nearly all genomic landscapes should...targeting with near-PAMless engineered CRISPR-Cas9 variants. Science 368,290-296(2020). DOI:10.1126/science.aba8853...PAM compatibility and editing activities of Cas9 variants. Nucleic Acids Res. 2021 Sep 7;49(15):8785-8795...Expanding Beyond Cas9 xCas9: Engineering a CRISPR Variant with PAM Flexibility History of CRISPR Cas – A ...

  2. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...unique tropisms are now available. These include variants with broad CNS or PNS transduction, cell type ...  Other model organisms Tropism and efficacy can vary depending on species. LY6A and LY6C1 do not have...Tabebordbar et al., 2021) method, selecting for variants that were effective in C57BL/6J mice, BALB/cJ ...transduces NeuN+ neurons, with transduction efficiency varying from 10-70% in different regions of the brain (... by transferring the 7-mer peptide region from variable region VIII of AAV9-X1.1 to that of AAV1. AAV1...BALB/cJ and C57BL/6 mice and rats. In mice, no variation in transduction efficiency was seen between different...lower than those seen with AAV9, they were highly variable and were not lower than those of AAV-PHP.eB (Goertsen...

  3. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...prepared buffer. This is because standard curves vary depending on a number of factors, including the ...equipment, assay incubation time, etc., which can vary even when the same protocol is being followed.  ... to calculate its experimental value. Though it varies from assay to assay, typically the experimental...trendline for the data. The appropriate trendline varies between assays and may be linear or nonlinear. ... of analysis. Different fields and assays have varying criteria for an acceptable R2 but for many scientific...allows you to calculate the curve’s coefficient of variation, or %CV. The %CV of a sample is the standard deviation...the %CV, the more accurate the curve. Though it varies based on the assay, the ideal %CV is less than ...

  4. Plasmids 101: Broad Host Range Plasmids

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    Blog Post
    ... different bacteria, and can be transferred to various experimental hosts (Jain and Srivastava 2013). ...structural elements that are versatile and adaptable. Variation in ori structure allows plasmids to utilize diverse...for replication initiation proteins. These iterons vary in their sequence and number and can alter the efficiency... their structural elements, oris interact with various RNA polymerases and proteins from bacterial hosts...proteins (RepA, RepB) necessary for replication in various hosts (Pagatto and Dillon 2001).  The plasmid has...broad host range plasmid with multiple oris that vary in DnaA-box and iteron structure, host-independent...the Broad Host Range Plasmid RK2: Positioning of Various Motifs is Critical for Initiation of Replication...

  5. Hot Plasmids - August 2020

    Type
    Blog Post
    ... can vary significantly depending on transgene design and delivery. To overcome these variations in human...utility of the system for use with low affinity variants of TEV. The new system has been demonstrated to...the evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription...fluorescence protein-based calcium sensors (jYCaMP1), variants of the calcium indicator jGCaMP7, that are optimized...new calcium sensors include syn-jYCaMP1s (slow variant), the Cre-dependent syn-FLEX-jYCaMP1s, and the ... Various Addgenies...

  6. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...reach all regions of the CNS. PHP.B, a neurotropic variant of AAV9, is better than AAV9 at gaining access ...PHP.B viruses. Creating PHP.B, a neurotropic AAV9 variant PHP.B’s specificity for C57BL/6J mice is likely...screened in vivo in C57BL/6J mice strains to identify variants that reached the CNS following IV injection. Compared...efficiency, but at a log lower dose than PHP.B.  A Ly6a variant is linked to PHP.B CNS permissiveness Both the ...permissiveness. The Wilson Lab also looked for the gene variant responsible for the PHP.B permissive phenotype ...how the method used to select novel AAV capsid variants could unintentionally limit the utility of candidate...E., et al. "Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain...

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

    Type
    Blog Post
    ...types or organisms, promoters must be similarly variable. Bacterial promoters only work in prokaryotic ...species from which they were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants...gene.  Constitutive  Widespread expression, but may vary by cell type. Tends to resist promoter down regulation...operators. Good for modulating gene expression through varied inducer concentrations.  Sp6 in vitro transcription...well-suited for modulating gene expression through varied inducer concentrations. trp High levels of gene...promoter. Good for modulating gene expression through varied inducer concentrations. Ptac General expression...regulation. Good for modulating gene expression through varied inducer concentrations. Generally better expression...

  8. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...neuronal activity. Researchers have designed a variety of photoswitchable elements into many other types...available to disrupt microtubules in cells, they have various disadvantages and limitations – for example, microtubule-destabilizing... Opto-katanin will surely be useful for a wide variety of experiments. Opto-RhoGEFs manipulate cell morphology...other GTPases. The Chow Lab has also generated a variety of membrane-recruitment optogenetic tools for manipulating...feasible or when the protein of interest supports a variety of processes in different cell types. One great...phospholipase D (PLD). Now they present a set of variants called SuperPLDs with 10, 30, 50, and 100 times... and cytotoxicity better than the 50x and 100x variants. But those super-high efficiency constructs are...

  9. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...are numerous GFP variants. Here I use the name ‘GFP’, which refers to mEGFP. This variant was made by introducing...jellyfish GFP (AvGFP) to create enhanced GFP (EGFP), a variant with improved brightness (Tsien, 1998). Another...necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al., 2002). GFP needs oxygen...Cranfill et al., 2016 and FPbase.org). A number of variants remain fluorescent under acidic conditions, with...can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can...although it still is worthwhile to generate several variants and check the functionality of the fusion (Mastop...

  10. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...recombinase variants to optimize PASSIGE efficiency (Pandey et al., 2024). The best-performing variant yielded...TALEs), with fifteen total variants deposited with Addgene. These variants, including a “miniCRISPR” version... linkers and spacers could enable many future variations using nanobodies or scFvs for other targets or...methyltransferase. The strategy is generalizable to a variety of DNA-targeting proteins, so the team created ...allowing the team to develop self-silencing CHARM variants that stop their own expression after silencing...

  11. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...owing to its ease of use and utility in a wide variety of organisms.  While much of current CRISPR research... in plants is also being realized. There are a variety of reasons to consider using genome editing to ...for the use of CRISPR in plants, and outline the various plant genome engineering tools that are available...systems, expression of S. pyogenes Cas9 or Cas9 variants (hereafter referred to as Cas9) and a single stranded... the nucleus, and several codon optimized Cas9 variants have been created in an effort to increase translation... Editing Kit from the Stuttmann lab contains a variety of Agrobacterium-compatible, Cas9 containing vectors...plant gene; you can find many plasmids for a wide variety of CRISPR applications in plants available through...

  12. 9 Tips to Achieve Success in Academia

    Type
    Blog Post
    ...London), Dr. Connie Cepko (Harvard Medical School), and Dr. George Church (Harvard Medical School) who took...We've interviewed Addgene depositors who are at various stages in their careers to get advice on choosing..., you're doomed.” 2. Always be sure to include various positive and negative controls to “convince [any...experiment is definitely not a fluke.” 3. Engage in various projects and help colleagues with their work. “...critical thinking skills that are applicable to a variety of careers outside of academia. She has had previous...

  13. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...circular plasmid that can be easily maintained in a variety of biological systems. Read on for an in-depth ...backbone. Ideally, the backbone will contain a variety of restriction enzyme cut sites (restriction sites...completion. The time required for complete digestion varies for different enzymes. Many companies now sell ...be cut out of the gel individually. There are a variety of ways to visualize the DNA in your gel (this ...approximately 1:3. Since the number of base pairs for each varies, it is difficult to calculate this based on DNA...for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert. It is critical... plating on the appropriate antibiotic and try varying the “recipient plasmid : insert ratio” in the ligation...

  14. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...was the Head of Oncology Target Discovery at the Novartis Institutes for Biomedical Research. Drug development... genetics.” As a postdoc in Stephen Elledge at Harvard Medical School, he developed the first lentiviral...are less common, they can be very impactful. At Novartis, Stegmeier deposited the ClonTracer Barcoding ...an Associate Professor at Mass Eye and Ear and Harvard Medical School (HMS) where he leads a lab of 25...also teaches at HMS and has been involved in a variety of commercialization efforts in gene therapy either... an Associate Member at the Broad Institute of Harvard and MIT. ...

  15. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...changes of these Twitch sensor variants were optimized by testing various linker configurations in a large-scale...from muscle cells. They identified a toadfish TnC variant which had a high-affinity for calcium binding and...large-scale functional screen in bacteria. Sensor variants were then refined by a secondary screen in rat...“trick” to the development of additional color variants. This resulted in the construction of redCALWY...redCALWY-1, a red-shifted FRET sensor for zinc using variants of mOrange2 and mCherry as donor and acceptor ...

  16. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...proteins, to study or alter their function for a variety of purposes. One focus of bioengineering is antibody...Antibodies can be broken up into smaller parts, the variable regions can be altered for specific functions,...IgG: immunoglobin G; ScFv: single-chain fragment variable; sdAb: single-domain antibody (often referred ...primers are a group of primers that contains a variety of degenerate codons (different codon sequences...sequences that code for the same amino acid) for various amino acids. Essentially, instead of forcing sequence...therapies and improve their delivery methods to various cells. The wealth of smaller engineered antibody...19px;font-family: helvetica;","color":"#FFFFFF","variant":null,"font":"helvetica","style":"font-family: ...

  17. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    ...biosensor tools, we now offer the jGCaMP7c variant in AAV1. This variant exhibits high contrast between its resting...serotype testing. These samples are available in various serotypes of two popular control AAV encoding GFP...basal fluorescence of the sensor. This “contrast” variant provides improved detection for tracking large ...-WPRE 105322  AAV1 pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE 111262  AAV1 pAAV-hSynapsin1-axon-GCaMP6s...Name 105322  AAV1  pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE 105714  AAV8  pAAV-Ef1a-fDIO-GCaMP6s...

  18. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ...Klebsiella spp. Dr. Devarshi Gajjar, The M. S. University of Baroda, India  Devarshi Gajjar has 10 years...antibiotic susceptibility of biofilms formed on various structured surfaces. Plasmids expressing fluorescent...astrobiology. During the last 11 years he has studied various aspects extreme life on earth including extremophile...scientists globally, Addgene has partnered with various community organizations and workforce projects ... Various Addgenies...

  19. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...be used to study dynamic processes. Conversely, varied input patterns may also be used to engineer new...response regulator (RR). TCS-induced responses are varied; one outcome is directed transcription from a given...obligate chromophore allows the system to respond to various light inputs. In far-red light or dark conditions.... A hybrid histidine kinase/photosensor senses various light inputs, and an obligate chromophore allows...They experimented with the promoter strength of various components, removing inducible promoters that could...many of these systems could be made suitable for various scientific or engineering applications.   References...

  20. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...technology. Nano-lanterns contain a Renilla luciferase variant fused to an FP; when supplied with a luciferase...collection of multicolored nano-lanterns for use in various applications, including optogenetics, biosensors...BRET). Photons emitted by a Renilla luciferase variant are used to excite the fused fluorescent protein...requires a linear unmixing algorithm to separate the various colors. In fluorescent imaging, you use the excitation... wavelength partially separates the signals of various Nano-lanterns, but it does not isolate each Nano-lantern...Addgene. Suzuki, Kazushi, et al. “Five colour variants of bright luminescent protein for real-time multicolour...
Showing: 161 - 180 of 770 results