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  1. Custom CRISPR Screens & the Green Listed Software

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    Blog Post
    ...the user. This approach is useful for several applications as will be discussed below. The software could...has a maximum of one gene knocked out. In this genetically heterogeneous population, cells behaving differently... you could majorly accelerate progression into clinical trials. Secondary screens  If you have generated...genes you’d like to test for phenotypic effects. Typically the phenotype would be studied in vitro, but there...contain multiple gRNAs targeting all genes in the indicated species and were suggested to be the best gRNAs...
  2. Reflections and Looking Ahead

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    Blog Post
    ...Antibody characterization is critical to enhance reproducibility in biomedical research. eLife, 13, e100211...tell only part of the story. This year, we made significant strides in making science more accessible and...launched AAV Packaged on Request, a service that dramatically expands and simplifies your viral vector options...
  3. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

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    Blog Post
    ...promoter. It can be used to drive Cre expression specifically in neurons. AAV Delivered Cre - A useful tool...Indeed, this AAV provides you with a means to specifically activate expression of any Cre-dependent gene...pmSyn1-EBFP-Cre Of course there are also more complicated ways to use this particular AAV. For instance...Additionally, you could use the retrograde form to specifically express Cre in neurons with axons that extend...
  4. Addgene is Expanding Our Viral Vector Service!

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    Blog Post
    ...service. A small subset of AAV plasmids will have technical or legal restrictions that make them ineligible...be reviewed by Addgene for legal approvals and technical compatibility with our production process. This...request is approved, you will receive an email notification that your request is ready to order. An “Add...found in our in-stock preps. If a request has technical considerations that may affect expected titers...
  5. Cre-ating New Methods for Site-specific Recombination in Drosophila

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    Blog Post
    ...incredibly useful molecular biology tool, but like any biological system, it has certain drawbacks. First, the ...Gerald Rubin’s lab sought to make complex genome modifications in Drosophila using multiple recombinases. To...offspring carried both the recombinase and GAL4, indicating that the recombinases were inherited normally...Chinese hamster ovary (CHO) cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin...
  6. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

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    Blog Post
    ...  “This enables you to do customized genetic modification on a scale that was really not possible before...genome engineering with CRISPRs was limited to modification of a few loci at a time,” Wang said. “It was... lentiviral libraries and their demonstrated application offer proof-of-principle that CRISPR-Cas9 can...aimed for 300 cells per construct, but in some biological systems one “might need larger numbers.” Further...
  7. Addgene's a Nonprofit? Nonprofit Awareness Day 2017

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    Blog Post
    ...service organization like the Red Cross or the American Cancer Society, or maybe you think about a local...services we provide like quality control, curation, technical service and open-access educational resources....decisions at Addgene If you’re ever curious what specifically drives the decision making process at Addgene...Gizela, Henry, Ina, Inna, Ishrat, Jane, Jason, Jessica, Joel, Jonathan, Justine, Karen, Kate, Kayla, Kelly...
  8. An Addgene Co-op: The Intersection of Fun, Friendship, and Work

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    Blog Post
    ...time for me to complete an online social media certification on social media data, and I was given a fantastic... occasional TikTok. I was also responsible for typical entry level tasks such as stuffing new depositor...connections with coworkers seem, in retrospect, nonsensical. I have found the people at Addgene have become... at Northeastern and Addgene co-op studying communications, marketing, and global fashion studies. She...
  9. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

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    Blog Post
    ...: Find sequences for plasmids assembled from publications and commercially available sources, as well ... as expression patterns, post-translational modifications and polymorphisms Codon optimization tool: To...making it ready to copy and paste into other applications Primer design tools Many of the above-listed... well as primer design for different cloning applications. Primer3 and Primer3Plus: For general primer...
  10. Addgene at the ASM Microbe 2016 Conference

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    Blog Post
    ...symposia on Sunday afternoon were dedicated to science communication through art. Addgene Advisory Board...page and, just a few weeks ago, we attended the American Society for Microbiology's annual meeting (ASM... Jessica Welch...
  11. Plasmids 101: Antibiotic Resistance Genes

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    Blog Post
    ... perspective – a plasmid-containing cell must replicate the plasmid in addition to its own chromosomal...How else can antibiotics be used in the lab? Historically, antibiotics have also been used to disrupt ...cloning. In these instances, the cloning vector typically has two separate resistance cassettes and your...and can be used in place of Ampicillin in most applications. Antibiotics vary in their sensitivitly to heat...
  12. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    Blog Post
    ... lab at ETH Zürich. “Instead of a graphical abstract, it’s a musical abstract,” says Di Ventura, a professor...in @nchembio Title: Bye bye L-arabinose drive. Dedicated to Edo, Armin and @KhammashLab. I hope you all...reached the level of expression needed for most applications.   Figure 1: With blue light, the VVD...
  13. Using Addgene's New Viral Service for Your Research

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    Blog Post
    ...preparations have worked well in my lab, including in applications where we have delivered them in vivo to the ...Roth’s lab are already available allowing you to chemically control neuronal activity while studying your...distributing new types of materials. At the time of publication we’ve already fulfilled more than 140 orders ... service as we continue to develop it. More communication and sharing will only help research advance....
  14. Visualizing Translation at the Single Molecule Level

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    Blog Post
    ...physiology. Ribosome profiling Ribosome profiling, a biochemical method developed in 2009, takes a “snapshot” ...translation While ribosome profiling co-opts the physical location of the ribosome, TRICK (Translating RNA...development, the osk-TRICK reporter was double labeled, indicating translational repression. During later stages...studying translation in various cell types and physiological states. References 1. Ingolia, Nicholas T.,...
  15. With Our New Viral Service, We're Taking CRISPR Further!

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    Blog Post
    ...represent the fraction of that total with the indicated function. One huge reason CRISPR has become such...Chavez, researcher at the Wyss Institute for Biologically Inspired Engineering and member of the Addgene...achieving the gRNA representation recommended in the publications associated with these libraries. Please note...particular, the Church lab at the Wyss Institute for Biologically Inspired Engineering and the Zhang and Doench...
  16. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

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    Blog Post
    ...microorganisms based on metagenomic data. MerMAIDs (Metagenomically discovered Marine, Anion-conducting and Intensely...publicly available metagenomic datasets with biophysical studies and the potential of mining datasets ...plasmids at Addgene Opperman J et al., Nature Communications, 2019. PubMed PMID: 31346176. Using LADL to...CIBN), which they call the LADL Anchor. Using specifically designed gRNAs, they can then recruit these ...
  17. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...ll need a few controls specific to flow as an application. These controls will allow you to distinguish...sample. The isotype antibody will be virtually identical to the antibody for your marker of interest. It...commercial antibody pages. Flow cytometry can be a complicated and daunting process. Controls like single color...research technician in Arlene Sharpe's lab at Harvard Medical School.   More resources on the Addgene blog Introduction...
  18. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

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    Blog Post
    ...study it. The process is fairly straightforward. Basically, you place your gene of interest in a plasmid ...genetic crosses with balancer chromosomes to make genetically stable fly lines. After a few weeks of waiting...throughout the lifespan of the fly to determine the implications of your genetic manipulations. You can produce... aware of at least some of the possibilities. Basically, if you can devise an experimental assay that ...
  19. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

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    Blog Post
    ...Immunosurveillance and Defense With increasingly sophisticated visualization techniques it is possible to follow... and correlating them with tumor mutations and clinical responses. This platform has the potential to ...the use of optogenetic tools is expanding to immunological research. Here the Zhou group presented Opto-CRAC... et al. "Caloric restriction mimetics enhance anticancer immunosurveillance." Cancer cell 30.1 (2016):...
  20. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    Blog Post
    ... light-mediated? The researchers split the proteolytically active light chain of BoNT/B into two fragments...that undergo light-induced dimerization. Upon application of blue light, iLID undergoes a conformational...rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue performance,...the field. Arlene Sharpe’s group at the Harvard Medical School developed a solution: CHIME, a CRISPR-Cas9...
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