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We narrowed to 704 results for: des.1

Showing: 241 - 260 of 704 results

  1. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

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    Blog Post
    ...’s fused to the C-terminus of DsRed.T3. See Table 1 for a summary of Rosella’s excitation and emission...fluorescence Rosella emits based on its localization. Table 1: Summary of Rosella’s excitation and emission spectra... variants are available from Addgene. References 1. Rosado, C., Mijaljica, D., Hatzinisiriou, I., Prescott...single fluorescent biosensor. Scientific Reports,5(1). doi:10.1038/srep12397. PubMed PMID: 26215030. PubMed... phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally PHO8 ...

  2. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ...total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e., 10 and 50 µL of vector at 1 x 1013 vg/mL titer... and 1 x 1012 vg*. Most systemic capsids are easy to produce in high titer, but higher doses (>1 x 1012...selecting a capsid, while Table 1 gives a brief overview of published descriptions of commonly used or emerging...before using it in an application.     Figure 1: AAV toolkit for targeted gene delivery to the brain...Capsids page to learn more about that collection. Table 1: Systemic capsid plasmids available from Addgene ...AAV9. At lower viral titers than AAV-PHP.B (e.g., 1 x 1011 vg/mouse), AAV-PHP.eB can transduce a greater...Diverse Brain Cells. Annual Review of Neuroscience, 45(1), 447–469. https://doi.org/10.1146/annurev-neuro-111020...

  3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ... to other technologies like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas... can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath and 42°C ...nearly single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification... Not mentioned in the paper, likely approximately 1-2 hours Equipment needed: Mini PCR machines by DIY-Bio...Sensitivity: – 110 femtomolar  Time: Not mentioned, likely ~1-2 hours Equipment needed: PCR machines/Heat blocks...flattening the curve, it has become evident that widespread testing is crucial in controlling this pandemic...one temperature. Two Cas12a-crRNA complexes are designed, each binding to one strand of the reverse-transcribed...

  4. Plasmids 101: Restriction Cloning

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    Blog Post
    ...well, so let’s go over the details step-by-step. 1. Digestion Set up restriction digests for your insert...recipient plasmid : insert ratio” of approximately 1:3. Since the number of base pairs for each varies,...cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent cells...at minimum, two transformations after a ligation: 1. Control Transformation containing the ligation mixture...result in few colonies on both plates (unsuccessful 1), in a vector alone plate with many more colonies ...single enzyme if your insert is flanked on both sides by that enzyme’s restriction sites, but the insert...the orientation of your insert. You may want to design a diagnostic digest for this purpose. Ideally, ...

  5. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

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    Blog Post
    ...plasmids. The Zhang lab, on the other hand, uses a 1 plasmid, single infection system. Genome wide mouse...X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F. Science. 2013 Dec 12. (PubMed...Wang’s library is designed to target a smaller set of genes and it includes more guides per gene. The Sabatini...Two new human lentiviral CRISPR library systems described in companion papers in Science in December were...Additionally, a concurrent Nature Biotechnology article describes the development of a mouse lentiviral CRISPR library...

  6. Multifunctional Neural Fibers: Modulation from Gut to Brain

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    Blog Post
    ...Fun fact: A 1 cm block of polymer can generate many meters of fibers!     Fig. 1 - Overview ... and brain neural circuits. Nature Biotechnology, 1–13. https://doi.org/10.1038/s41587-023-01833-5 Resources...blocks of polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics...whistles’ – including microscale light emitting diodes, thermal sensors, and microfluidic channels. These...using NeuroStack and the fibers as previously described, then measured electrophysiological potential ...

  7. AAVs in Retinal Gene Therapy

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    Blog Post
    ...and Therapeutic Purposes. Annu Rev Virol. 2014 Nov;1(1):427-51. DOI: 10.1146/annurev-virology-031413-085355...personalized medicine. References and further reading 1. FDA approves gene therapy for blindness 2. Gene Therapy... Med Biol. 2014; 801: 687–693.  doi:  10.1007/978-1-4614-3209-8_86. PubMed PMID: 24664759. PubMed Central...whole field to a grinding stop and only now, 2 decades later, is it finally back in the limelight. On ...(such as viral vectors) without triggering a destructive inflammatory immune response. At the time, adenoviral...follow in the footsteps of Luxturna™. However, despite promising results in preclinical studies some potential...technology: engineering of new capsid variants, designing expression cassettes to regulate and optimize ...

  8. Expanded Access to Rabies Plasmids

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    Blog Post
    ... export license applications for VSV and SARS-COV-1 plasmids. Export control license process    An export...charged.  Timelines  We have offered VSV and SARS-COV-1 plasmids under this application process since May ...institution. Requesters will need to provide a brief description of the applicable research project when purchasing...

  9. A Look at Addgene's QC Process

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    Blog Post
    ...If a depositor provides a sequence for the plasmid, QC scientists will first align the de novo NGS result...experiments, to false conclusions.  Figure 1. Potential points where DNA errors can be introduced...are assembled into a circularized plasmid sequence de novo (without the use of a reference sequence) using...sequence, if one was provided. If no reference is given, de novo assembly in Geneious can still provide additional...plasmids in our collection. This standardization provides security for requestors, ensuring that the plasmid...types of data for each of their plasmids, from descriptions of key components of the plasmids to partial...believe may affect the function of the plasmid. This includes point mutations, frame shifts, missing elements...

  10. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...NGS) to identify genes of interest. Figure 1: Overview of in vitro AAV CRISPR screens. Created ...certain experiments. AAV can be safely handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires...CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo,...diseases. Signal Transduction and Targeted Therapy, 9(1). https://doi.org/10.1038/s41392-024-01780-w Additional... This vastly increases the possibilities when designing screens. Each serotype has its own tropism, or...experiment. Additional considerations One of the downsides to using AAV is the limited packaging capacity...Adames, A. C., Saur, D., & Rad, R. (2022). Tutorial: design and execution of CRISPR in vivo screens. Nature...

  11. The Materials Science of Optogenetics Experiments

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    Blog Post
    ... and materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various ...follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of motor...manipulation of neural circuits. Nature protocols. 2012;7(1):12-23. doi: 10.1038/nprot.2011.413. PubMed PMID: ...freely moving mice. Neuroscience research. 2011;70(1):124-7. doi: 10.1016/j.neures.2011.01.007. PubMed ...

  12. Visualizing Protein Turnover In Situ

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    Blog Post
    ...unpublished, that allowed the visualization of specific de novo synthesized proteins in situ. I was privileged..., 2010) and allows the visualization of the whole de novo synthesized proteome. However, FUNCAT alone ... contact her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization...radioactive materials is highly regulated, requires designated work areas and come with health risks. Importantly...synthesized endogenous proteins in situ. Now FUNCAT-PLA provides both quantitative and spatio-temporal information...

  13. Isolating Genomic Regions of Interest with the CRISPR System

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    Blog Post
    ...identification of proteins associated with the IRF-1 locus. Figure 1: Schematic of enCHIP. Image from Fujita...the promoter region of the transcription factor IRF-1. Immunoprecipitation, using an antibody targeting ...References Campbell, Amy E., et al. "NuRD and CAF-1-mediated silencing of the D4Z4 array is modulated ...region In 2013, the Hodaka Fujii Laboratory first described a method to purify a specific genomic region using...plasmid. Experimental use Since the initial report describing CRISPR-mediated purification of a specific genomic...

  14. DNA Purification Without a Kit

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    Blog Post
    ...Figure 1: Purify DNA using glass filters. Image: Kim and Morrison, 2009. This method provides DNA yield...a gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this syringe-based method... solution. NaI has two purposes in this protocol: 1) it solubilizes both DNA and agarose, and 2) it helps...property and developed a protocol that uses Whatman No. 1 paper or even paper towels to purify DNA and RNA from...plasmid purification and DNA gel extraction method described in Kim and Morrison is nearly identical to that...

  15. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...lentiviral gRNA expression vector (pRDA_526) that (1) enables efficient knockout (KO) of genes in immune...statistical power of in vivo screens. Figure 1: Using FITS to perform in vivo CRISPR screens to assess...single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation...assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission...also reveal novel pathways. The FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction...

  16. Quick Guide to Near-Infrared Fluorescent Proteins

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    Blog Post
    ...2021).   Table 1: Properties of modern NIR FPs     NIR FP Ex, nm Em, nm ECa, M-1cm-1 QYb, %   Molecular...expression level. State-of-the-art FPs (listed in Table 1) were optimized for efficient binding of endogenous...previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric state of a probe leads...iRFPs in cellular and molecular brightness (Table 1).  NIR reporters and biosensors NIR reporters can ...Biol 24, 758-766 e753 (2017). Baloban, M. et al. Designing brighter near-infrared fluorescent proteins: insights...

  17. Hot Plasmids: Summer 2024

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    Blog Post
    ...10.1038/s41551-024-01227-1. doi:  https://doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic...spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the purification and...highly-visible beads on the cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking... A) Schematic of workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single ...cryo-EM samples from damage at the AWI. Plus, tardigrades!  Find RvLEAMshort here!    Abe, K. M., & Lim...

  18. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...the viral TF library, with each cell receiving only 1 TF expression construct. Cells were cultured following...states was generated (Joung et al., 2023). Fig. 1: Barcoded TF library was packaged in lentiviral particles...atlas of directed differentiation. Cell. 2023. 186 (1) 209-229. doi: 10.1016/j.cell.2022.11.026 Joung J....transcription factor library The human genome encodes approximately 1,800 TFs, with >3,500 unique isoforms... and screening mechanisms, although the unique barcodes for each TF help streamline identification. While...a disease in a model system. TF screens can be designed to identify custom TF programs to acutely initiate...

  19. Antibodies 101: ChIP

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    Blog Post
    ...reserve a small aliquot, split into two parts to (1) run a DNA gel to confirm fragment size and (2)  incubate...with a phenol-chloroform extraction.    Fig. 1: Schematic overview of ChIP. Image courtesy of Shengliu... let’s do a quick review. At this point you have (1) crosslinked your proteins of interest to any DNA ...immunoprecipitation (ChIP) is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works...up the DNA, or enzymatic digestion. Sonication provides a truly random digestion, but is harsher and requires...both tissue or cell type, amount of protein, and desired fragment length for your output assay. A longer...

  20. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    ...dual-color control of neurons. Nature Communications, 12(1), 1–20. https://doi.org/10.1038/s41467-021-24759-5 Zhang...to consider when choosing one.     Figure 1: Example of an experimental optogenetic setup. A light... 21–51). Springer US. https://doi.org/10.1007/978-1-0716-0755-8_2 Lehtinen, K., Nokia, M. S., & Takala... 21–51). Springer US. https://doi.org/10.1007/978-1-0716-0755-8_2 Boyden, E. S., Zhang, F., Bamberg, E...In 2005, Boyden et al. described the first light-activated tool for controlling neuronal activity, channelrhodopsin... light in combination with retinal) variations designed to give researchers flexibility in using optogenetic...precise timing of the perturbation. Other tools are designed to be activated within a narrower band of wavelengths...
Showing: 241 - 260 of 704 results