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We narrowed to 302 results for: human dat

Showing: 261 - 280 of 302 results

  1. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...serotype 2-derived inverted terminal repeat sequences. Human gene therapy methods, 23 1, 18-28. PubMed PMID: ... virus for my first experiment. It was a bit intimidating, but I had my lab’s protocols and some great...hours hands-on time and 2 hours for the qPCR run and data analysis. Refer to this Addgene AAV Titration by...

  2. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...not fully suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi....resistance to both carb and amp, due to their common degradation mechanism. How to choose between the two? Carb...high pH environments and is less susceptible to degradation in agar plates. This prevents satellite colonies...

  3. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ...building events like Escape the Room or Habitat for Humanity take you and your co-workers out of the office... up and running repository. As they built the foundation for what Addgene is today, there was a lot of...culture here at Addgene has remained paramount. Candidates who interview at Addgene frequently ask the question...

  4. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...Multicolor CRISPR labeling of chromosomal loci in human cells.” Proc. Natl. Acad. Sci. USA 112(10) (2015...protein. Such flexibility makes aptamers ideal candidates for biosensors. Aptamer biosensors are used in.... The nature of aptamers makes them excellent candidates for developing assays for otherwise difficult-to-study...

  5. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...cytoplasmic environments. However, approximately 40% of human (and most eukaryotic) proteins localize to chemically...suggest starting by using the newly published oxidative optimized palette of FPs (moxFPs) [5]. The moxFPs...of cysteine-free fluorescent proteins for the oxidative environment." PloS one 7.5 (2012): e37551. Pubmed...

  6. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ... of transcytosis and transduction using primary human brain endothelial cells. J Neurochem 140, 216–230...Aebischer, P. (2010). Efficient transduction of non-human primate motor neurons after intramuscular delivery...endocytosed material or molecules targeted for degradation. These two pathways also use different cytoskeletal...comprised of the viral proteins VP1, VP2, and VP3. To date several hundred variants, or serotypes, have been...

  7. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and sgRNA expression ...previously validated gRNAs at Addgene. In vitro screening for the best gRNA Figure 1: Validation of double... a simple method for selecting the guide RNA, validating its efficacy in vitro, and using it in mouse ... protein (EGFP) reconstitution. (a) Scheme of validation for DSB mediated EGFP expression cassette reconstitution...vitro screening is a simple and fast method for validating your selected gRNA’s, and has the bonus benefit...benefit of supplying you with primers for validating your edit in mice once they have been generated (more...more on that later). Find Validated gRNAs to Target Mouse Genes To begin the in vitro screening process...

  8. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-secreting beta-like...usually hypothesized to be a result of chromophore oxidation. Some of the more popular timers (whether as part...

  9. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ... labelled with a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: ...instrument used in this reconstruction lay the foundation for all subsequent versions of LSFM microscopes...

  10. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...upregulates expression of IGF2, a characteristic of many human malignancies. Additionally, the Tapscott Lab identified...method allows for the recovery of a vast amount of data from a single experimental setup. As we’ve seen ...

  11. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ... in medicine, 27% in natural sciences, 82% in humanities, and 32% in the social sciences go uncited. Creative...the Evolution of Scientific Impact Data Freedom: The Expansion of Data Sharing in Research Publications ...example, learn about how researchers are using “big data” to look at Lyme Disease in new ways. Or learn about...at a time – A video by LeeAnne Thayer, a Ph.D. candidate in marine sciences at the University of Maine,...

  12. Adenovirus Guide

    Type
    Guide
    .... More than 80% of the human population has been exposed to at least one human AdV serotype and has developed...cells. AdVs can cause a range of mild diseases in humans and other animals, most commonly respiratory infections...expressed by the packaging cell line, normally the human embryonic kidney 293 (HEK293) cell line. There is...More than 60 serotypes have been identified within human adenoviruses, which are classified into seven species... vectors, although replication incompetent, are human pathogens and have to be handled at biosafety level...adenoviral vectors as vaccines for outbreak pathogens . Human Vaccines & Immunotherapeutics, 13 (12), 3020–3032...the left ITR, resulting in the possibility of accommodating ~36 kb of genetic cargo. Production of HCAdV...

  13. CRISPR Guide

    Type
    Guide
    ...) limiting. This PAM is abundant throughout the human genome, but an NGG sequence may not be positioned...activate or repress gene expression in mice and human cells. The simplest dCas9-based activators and repressors...CRISPRoff maintains gene silencing across much of the human genome following transient expression of the dCas9... mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology , 31 (9), 822–826. PMID...Norville, J. E., & Church, G. M. (2013). RNA-Guided Human Genome Engineering via CAS9. Science , 339 (6121...Yamamoto, T. (2014). Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system...Doudna, J. A. (2014). Enhanced homology-directed human genome engineering by controlled timing of CRISPR...

  14. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Editing in Human Cells Resources on Addgene.org Browse All CRISPR Plasmids Find Validated gRNAs for...Mary Gearing on Mar 12, 2015; updated by Christina Mork, Jul 27, 2020; updated by Susanna Stroik January ...

  15. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...genetic screening. gRNAs targeting every gene in the human genome for example, can be made easily and cheaply...Last updated Oct 7, 2020 by Gabrielle Clouse. This post was contributed by guest bloggers Marcelle Tuttle...through the creation of innovative technologies. To date his group has develop several popular CRISPR-based...

  16. Molecular Biology Reference

    Type
    Guide
    ...thousand basepairs from the entire human genome. Digesting the human genome with restriction enzymes would...plasmid in intended for use in human cells, the promoter will be a human or mammalian promoter sequence...molecules. In order to isolate a particular gene from human chromosomal DNA, it would be necessary to isolate...including plants, worms, mice, and even cultured human cells. Although plasmids were originally used to...strains of E. coli , some of which are deadly to humans. The majority of all common, commercial lab strains...concentrations. We suggest checking your plasmid's datasheet or the plasmid map to confirm which antibiotic...sequencing platform that can generate sequencing data for up to 600 billion bases in one reaction. In ...

  17. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...expression atlas of miRNAs and their promoters in human and mouse. Nature Biotechnology, 35(9), 872–878....methods are often quicker and the repositories of validated small RNAs are vast and difficult to beat in terms...knockdown cells lines, and you can usually find validated shRNA sequences for your intended target. RNAi...(2021). Life of RISC: Formation, action, and degradation of RNA-induced silencing complex. Molecular Cell...

  18. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ... al. "Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system." Cell 155.7... in fusion proteins. Since GFP is such a well-validated, all-round good performing probe, it is the first...components. For a review see Lotze et al. (2016). To date, small probes that are entirely genetically encoded...

  19. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...He has also constructed a library of eGFP-tagged human ORF clones to allow testing and comparison of different...how they can be used, and how they’ve been used to date . If you have any questions, please write to us ...time, Christopher Marx's lab has engineered and validated a new method called FREQ-Seq. This strategy allows...

  20. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...W, Khalili K. Elimination of HIV-1 Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing. Sci...approaches, and each of these gRNAs will need to be validated for high on-target binding and low off-target ...Resources onf Addgene.org Check out Addgene’s newly updated CRISPR Guide! Find CRISPR Plasmids for Your Research...
Showing: 261 - 280 of 302 results