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Showing: 261 - 280 of 1082 results
  1. Viral Vectors 101: Systemic Capsids

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    Blog Post
    ...transgene expression-induced toxicity in NHPs (Hordeaux et al, 2020; Hudry et al, 2023; Hinderer et al, 2018).... 2023 * indicates viral preps available. Mechanisms of crossing the BBB are indicated in parentheses..., the suitability of a systemic capsid for an individual application is dependent on a number of factors...like CREATE (Deverman et al., 2016), M-CREATE (Ravindra Kumar et al., 2020), iTransduce (Hanlon et al.... vg total in mice) may increase the chances of inducing an immune response against the capsids or transgenes... CLOVER repository of published systemic AAVs to find commonly used vector cargo designs and administration...deposited by the Maguire lab. In mice, it is sex-independent and does not show significant differences in ...
  2. Three Tips to Organize Your Lab Notebooks in the New Year

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    Blog Post
    ...handwriting or trying to find to where the binder was moved. You’ll have peace of mind knowing the information... and post-its to make finding your most used protocols & references easy to find It’s happened to all ...you can’t find the molecular cloning guide you were sure was in one of those four white binders. Thirty ...on the spine. For binders of protocols and data, consider using color coded binders and tabs (for different...Start by gathering that pile of lab notebooks and binder on your desk. Arrange them in chronological order...tasks, set deadlines, and arrange automatic email reminders. These programs can be synced to your online calendar...to other notes, and use the search bar to quickly find what you’re looking for. Tip 3: Share protocols ...
  3. Hot Plasmids and Viral Preps - March 2021

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    ...it easier for you to find and use the plasmids you need. Here's what you'll find in this post: Epigenetic...the auxin inducible degron (AID) system in C. elegans By Alyssa Cecchetelli The auxin inducible degron (... a light-inducible CRISPR system developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target...assembly Tracking cell-cell interactions New auxin inducible degron system for C. elegans New CRISPR plasmids..., 2021).   Schematic showing the auxin-inducible degron system. Image from Ashley et al., 2021....spatiotemporal expression pattern and an auxin inducible degron (AID) sequence attached to your protein...complex. In the presence of auxin, this TIR1 complex binds to the degron sequence ultimately leading to the...
  4. Hot Plasmids: Winter 2025

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    ... our Hot Plasmids articles.  Here's what you'll find in this post: Optimized prime editing in dicots...single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors kit Antibodies for neurodegeneration...potential to improve rapid and precise plant breeding. Find dicot prime editing plasmids here! Vu, T. V., Nguyen...kinesins but for many other proteins of interest too! Find CLIP, CLAP, and SNAP plasmids here! Smith, E.R....2024.12.20.629623.  A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic...and more responsive genetically-encoded voltage indicator (GEVI) than previous versions. ASAP5 can be imaged...The initiation site identified in panel (C) is indicated; red line is exponential fit. Adapted with permission...
  5. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    ... Addgene blog Find our Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips...Addgene.org Find fluorescent proteins for your research Read our Molecular Biology Reference Find plasmids...phytoplankton, and colorful corals, to name a few. What’s behind these sights are an abundance of biological molecules...limited or for bioart and teaching applications. Find a chromoprotein for your experiment! Eukaryotic ...contain a toxin gene (regulated by a rhamnose-inducible promoter) and a chromoprotein gene. Allelic exchange...expressed as it is under the control of a rhamnose-inducible promoter. The resolution step leads to the desired... backbone. To do this, rhamnose is added which induces expression of the toxin. Any cells that grow after...
  6. Cloning Mammalian Cells with the Agarose Method

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    ...blogger Iris Lindberg, Professor at the University of Maryland School of Medicine. In the Lindberg Lab we ...blogger Iris Lindberg, Professor at the University of Maryland School of Medicine. Iris Lindberg is a professor...grease, place and fill a dozen cloning rings, the remainder of the colonies on the plate dried out and died... cell clones with sterile truncated pipet tips. Indeed, an experienced operator can pick 48 clones from...mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...antibiotic-resistant colonies, use a Sharpie to circle 12-48 individual colonies by (carefully) holding plates up to ...identified from the plate circling- are then individually punched out, gently sucked up into the pipettor...
  7. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

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    ... DNA-binding molecules for the first time and named the new technology as engineered DNA-binding molecule-mediated...also developed the inducible translocation trap (ITT) system to analyze signal-induced nuclear translocation...immunoprecipitation (iChIP) and engineered DNA-binding molecule-mediated chromatin immunoprecipitation...insertion of recognition sequences of an exogenous DNA-binding protein such as LexA into the target locus, which...is the rate-limiting step. When engineered DNA-binding molecule technologies such as TAL proteins and ...molecules makes it much easier to knock-in LexA binding elements into the target region. However, it struck...struck me that direct use of such engineered DNA-binding molecules even enables us to skip the step of knocking-in...
  8. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

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    ... fluorescent protein guide Find plasmids for subcellular localization Find fluorescent protein plasmids...Part stable scaffold, part epitope binding region, all fused to a fluorescent protein. “Frankenbody,”...antibodies for tagging purposes. Different antibodies bind different protein sequences called epitopes and ...interest. Antibody based probes, such as antigen-binding fragments (Fabs) (Porter 1959) and single-chain... 1972) are fused to mature fluorophores, and can bind and light up epitope tagged proteins in vivo. These...CDRs, or loops) of an antibody that recognize and bind specific epitopes (Ewert et al., 2004). This scFv...to easily observe the tagged protein upon probe binding in living cells.  In their proof of concept experiment...
  9. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

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    ...stained with PI (pink) to indicate cell death. Image from Diessl et al., 2020. Find the GFP reporter calcineurin...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like... for a low number of cells and in vivo settings. Find the sgRNAiBAR library!  Zhu et al., Genome Biology...cells in a population, it’s easier to visualize individual/non-overlapping cells. Until now, tools for doing...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ...dephosphorylates the protein CZI, which in turn binds to calcineurin-dependent response elements (CDREs...fusions with tTALE to “stain” DNA. TALE proteins can bind specific DNA sequences which brings the attached...
  10. Troubleshooting and Optimizing a Western Blot

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    ...Antibody not completely binding Increase incubation time Not enough antibody binding sites Polyclonal...the technical design of your blot, but are still finding yourself having issues, you’re in the right place...compartment. If your proteins are nuclear- or DNA-binding proteins, you may need to sonicate your lysates...lysates in order to release the proteins from their binding. If you’re seeing protein aggregation, you may ...or improperly made running buffers can cause all kinds of odd issues. If your gel is running properly but...ladders contain either ‌a protein tag or an IgG binding site, allowing you to visualize them with a conjugated...double-membrane transfer with a psi membrane stacked behind the p membrane. If your protein runs through the...
  11. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    ...editing or the induction of iPSCs, are available in the Resource Center. Researchers can also find links to ...so that both veteran and new Rett researchers can find the reagents and resources they need to cure Rett...Syndrome? Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome is a rare...normal development through ages 6 to 18 months, individuals with Rett syndrome experience regression of previously...Syndrome (Amir et al., 1999). We now know that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides...excision of the Stop cassette using tamoxifen-inducible Cre not only halted the neurological damage but...regardless of age or symptom severity. This exciting finding injected hope that a similar reversal may be possible...
  12. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    ...). The 3-5 nucleotide PAM sequence serves as a binding signal for Cas9 and this sequence is a strict requirement...identified four novel SpCas9 variants with altered PAM binding specificity: SpCas9 variant Mutations (relative...VRER variants is similar to wild-type SpCas9, indicating that the variants are likely just as selective...sequence may not be characterized Non-SpCas9's bind a variety of PAM sequences, which makes them useful...550:280–284 . https://doi.org/10.1038/nature24049 Find plasmids from this paper at Addgene. Amrani N, ...–1027 . https://doi.org/10.1126/science.aaq0180 Find plasmids from this paper at Addgene. Kim D, Kim...Biotechnol 34:863–868 . https://doi.org/10.1038/nbt.3609 Find plasmids from this publication at Addgene. Kim ...
  13. CRISPR-based Genome Editing Systems For Editing in E. coli

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    ... self-curing, an arabinose inducible lambda-red gene, and an IPTG-inducible sgRNA sequence targeting the...from our CRISPR topic overview Find other CRISPR methods for bacteria Find all articles about microbiology...editing, pTargetF can be cured from the cell by inducing expression of the sgRNA that targets the pMB1 ...updated pEcCas/pEcgRNA system, and confirmed that indeed the gRNA-pMB1 had a slightly higher leaky expression... includes versions of plasmids with different induction systems, such as PT7, Prha, Ptet, and Para. The...that is 2.6 times higher when compared to the industrial strain. In our metabolic engineering case study...high-yielding strain which had 6 times the yield of the industrial strain. Not only is MUCICAT better, it is faster...
  14. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    ...possible to reprogram differentiated cells into induced pluripotent stem cells (iPSCs) by the addition ...perform cell therapy and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources of adult...and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two sequences are necessary...cells [8]. The EBNA-1 protein recognizes OriP and induces amplification of the episomes concurrently with...such low efficiency of reprogramming is a major hindrance in using PBMCs as a source of somatic cells for...Cukier, and Derek M. Dykxhoorn. "Blood Derived Induced Pluripotent Stem Cells (iPSCs): Benefits, Challenges...PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced Pluripotent Stem Cells from Blood Cells: An Efficient...
  15. Tips for CRISPR Gene Editing in Mice

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    ... Resources on Addgene.org Find Software Tools to Help Design You gRNA Find CRISPR Plasmids for Use in... probably got a gene you’d like to manipulate in mind. Now it’s time to consider where you would like ...DNA supply company you currently use. You can also find many previously validated gRNAs at Addgene. In vitro...control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted from Mashiko...once they have been generated (more on that later). Find Validated gRNAs to Target Mouse Genes To begin the...plasmid using standard cloning techniques (The XX indicates where your target is inserted, disrupting the ...your modified pCAG-EGXXFP plasmid along with the individual gRNAs in their PX330 plasmids (or similar plasmids...
  16. Hot Plasmids - January 2023

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    ...benefit from region-specific, inducible expression.   Find heat-inducible lentiviral plasmids here!   Wu...what you'll find in this post: CRISPR-Csm: A novel RNA targeting system Heat-inducible lentiviral Car...Adapted from Figure 3 Colognori et al 2022.   Find CRISPR Csm plasmids here! Colognori D, Trinidad ...Epub ahead of print. PMID: 36690762.   Heat-inducible lentiviral CAR constructs by: Brian O'Neill  A...lab.  The crux of the system is a set of heat-inducible lentiviral CAR constructs, the most basic of which... which is depicted here: The heat-inducible Hsp (heatshock promoter) drives expression of the tumor-... signal in immunoassays due to only having one binding site per protein. Having multiple antibodies allows...
  17. Visualizing Translation at the Single Molecule Level

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    ...ribosome moves along an mRNA, it displaces other RNA-binding proteins to allow translation to occur. For a TRICK...requires the following components: a GFP protein that binds to the coding region of a reporter mRNA via a PP7... coat protein (NLS-PCP-GFP) an RFP protein that binds to the 3’ UTR of the reporter mRNA via an MS2 coat... the fluorescent proteins and is driven by an inducible promoter (Plasmid 64542 or 64543) Before translation...signal, NLS-PCP-GFP then returns to the nucleus to bind cognate untranslated mRNAs. The high specificity...multiple copies of the PP7 and MS2 coat protein binding sites to tether many copies of the FP-coat protein...development, the osk-TRICK reporter was double labeled, indicating translational repression. During later stages...
  18. Technical Design of a Western Blot

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    ...assay run time to two days.  Indirect or direct detection method Indirect detection is the most common...quicker to use, with high binding capacity, though it has lower retention after binding and washes. It’s less...for validation data, or follow the references to find the original publication and ensure that it includes...for a western blot. Polyclonal antibodies, which bind to multiple sites on an antigen, are cheap and can...the manufacturer, each rAb in a mixture can be individually validated. ‌rAbs are not as commonly available...). Direct westerns are simpler and faster than indirect ones and can be used in highly quantitative assays...tested to ensure it doesn’t impact the antibody’s binding to your protein of interest. Protein preparation...
  19. What's New in CRISPR - Winter 2018

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    ...factor localization and binding via ChiP-seq. Learn more about ENCODE Find the transcription factor...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find new CRISPR tools for your research.     Improved...sensitivity, and more. Read the paper in Science Find the new SHERLOCK plasmids More precision with ...Cas9’s. Read the paper in Nature Communications Find the Cas9-Cas9 chimera plasmids A modular toolkit...engineering in plants. Read the paper in bioRxiv Find the plant genome editing toolkit New Cas protein... Addgene blog post Read the paper in Science Find the Cas14 plasmids Anti-CRISPR proteins for optogenetic...
  20. Special Delivery: Fluorophore Targeting for FRET Studies

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    ...substrate or add an essential cofactor? How many binding partners does it have and how do they come off ...package needs to go to the proper mailbox (i.e., a binding site) in the right city (protein). In a cell, this...thousands of biomolecules each containing multiple binding crevices that can nonspecifically soak up your ...when donor and acceptor are close together. The binding of acceptors >100 Å away from the donor doesn’t...translation, the novel tRNA/unnatural amino acid finds the UAG codon in the ribosome, resulting in site-specific... reagents You may already have a FRET acceptor-binding site on your protein of interest: you just don’...NTA-sepharose beads used for protein purification: the NTA binds to the His tag on your protein and the coupled Cy...
Showing: 261 - 280 of 1082 results