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  1. Plasmids 101: Gateway Cloning

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    Blog Post
    ... The entry clone and destination vector carry different antibiotic resistance markers (indicated here ...STEP 1: Generate an Entry Clone There are a few different ways to generate our desired entry clone - human...interest: Find Gateway cloning plasmids here! References 1. Chee JY, Chin CF (2015) Gateway Cloning Technology... Digest Video Check Out Our Molecular Biology Reference Pages  ...
  2. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ...forward for what the field can do.    References and Resources References  Özcan A, Krajeski R, Ioannidi E,...RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using Cas7-11   Use ...fibrosis, which can be caused by thousands of different mutations, all found in the same gene. If we only...
  3. Finding nucleic acids with SHERLOCK and DETECTR

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    Blog Post
    ... studies.     Jennifer Tsang contributed to updating this article. References Abudayyeh OO, Gootenberg...variations of Cas13 and Cas12a, are combined with different fluorescent reporters for each enzyme. In addition...Zetsche et al., 2015 ). Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends...
  4. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, packaging...equally potent Cas9 protein, the Zhang lab took a different approach. They analyzed over 600 Cas9 orthologs...Sontheimer lab used NmeCas9 with AAV to edit two different disease-causing loci in mouse liver.  Two other...have also been used to express base editors. References Chew WL, Tabebordbar M, Cheng JKW, Mali P, Wu...
  5. Designing Your Chalk Talk for the Academic Job Interview

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    Blog Post
    ...if your model is correct or your data favor a different model. Finding evidence for or against a model.../goal, an approach, anticipated outcomes, and preferably a description of the payoff if the aim is successful...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...
  6. Hot Plasmids Spring 2024

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    Blog Post
    ... optical pooled screens in human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin...editing efficiencies in a number of cell types at different genomic loci — in some cases, over 20-fold higher...prime editor. B, Prime editing efficiencies at different loci in U2OS cells. Image adapted from Yan et ...that an A122D mutation in the rhodopsin domain conferred several new improvements. The new variant has ...
  7. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

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    Blog Post
    ...options in our CRISPR 101 series!  References and resources References  Anzalone, A. V., Koblan, L. W. &...plasmids that have been optimized for expression in different species and cell types including (but not limited...101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled Library...
  8. Using Video to Share Your Science: We Share Science

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    Blog Post
    ...that “… the emergence of the video abstract genre offers a low-barrier opportunity for researchers to leverage...to measure the impact of their contribution. It offers a 5-step process and several examples for creating...professors at the University of Washington in St. Louis, offer guidance on how to make a low-cost whiteboard video...WeShareScience or email Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric...
  9. 5 Reasons to Use Reddit for Science Communication

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    Blog Post
    ...of conversations with people who work in many different scientific fields. They also host AMA forums where...team with a description of our AMA and Joanne’s preferred time to start answering questions. r/Science has...AMA is now a great resource for any scientists to refer to if they have questions about science careers,...future. The AMA was also cataloged for permanent reference by the Winnower. The only small downside to the...
  10. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    Blog Post
    ...Vader in "Return of the Jedi" While Vader was referring to the evil side of a mystical "Force," this quote...cannot be ignored. They have the potential to interfere with the function of resident ER chaperones and...University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...Kaposi’s sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent...
  11. Data Freedom: The Expansion of Data Sharing in Research Publications

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    Blog Post
    ...of veracity of the conclusions drawn. The main difference with the PLOS policy is that access of this data...perhaps similarly impoverished lab) might apply different or novel tools and assumptions to reveal findings...geneticists may track the motion of multiple animals on different occasions via video, generating huge amounts of...responsible for storage.  If datasets are too large to transfer to a journal repository, they are likely too large...
  12. Adeno-associated Viruses (AAVs) for Genome Editing

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    Blog Post
    ...flanking a FLOXed neoR gene. pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR...Schofield, 2015; Moser and Hirsch, 2016; Epstein and Schaffer, 2017) Hopefully you’ve been able to check out...cohesin gene inactivation in human cancer.     References 1. Russell, David W., and Roll K. Hirata. "Human...27280971. 10. Epstein, Benjamin E., and David V. Schaffer. "Combining engineered nucleases with adeno-associated...
  13. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    Blog Post
    ...of interest supports a variety of processes in different cell types. One great way to do this is to use...SuperPLD module is optogenetically recruited by different CIB constructs to the plasma membrane or lysosomes...future updates to the collection! Resources and references Additional resources on the Addgene blog Viral....org Optogenetics Guide Optogenetics Plasmids References Aljiboury, A. A., Ingram, E., Krishnan, N., Ononiwu...
  14. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

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    Blog Post
    ...nucleotide insertions, dele­tions or other mutations, referred to as ‘indels’. While this is convenient for generating... iPSCs (or correct them back to wild type), differentiate them into somatic cells affected by the disease...iPSC clone picking, a 100-fold decrease is the difference between picking 100 versus 10,000 clones! We ...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering...
  15. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...to a mouse model of muscular dystrophy. Three different splice site donor and acceptor sequences were ...reconstituted after integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains...greater and more consistent transgene expression. References: Allocca M, Doria M, Petrillo M, Colella P, Garcia-Hoyos...retinal gene therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out ...
  16. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    Blog Post
    ...insertions, like a fluorescent protein knock-in, suffer from low efficiency when a standard dsDNA repair... secondary structure. Today, multiple companies offer long single-stranded DNA production services or ...Quadros et al. attribute much of this variation to differences in guide RNA efficiency. For example, in a two-guide...Quadros for assistance with editing this blog post. References Yang, Hui, et al. "One-step generation of mice...
  17. Generating Mouse Models Using CRISPR/Cas9

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    Blog Post
    ...directly into fertilized mouse eggs, circumventing the need for enrichment and selection offered by mouse ...into either the pronucleus or the cytoplasm of fertilized mouse eggs. Or, to avoid handling embryos ex ...therapeutic applications that better human health.     References Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib...
  18. A History of Genome Engineering in Popular Culture

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    ...TALENs more specificity and a broader substrate preference than the ZFNs.   Genetic Engineering Era: Post...genetically engineered superhuman Khan. The novel, Inferno (2013) and the movie version (2016) stars Tom Hanks...that are typically associated with the DSBs and offered unprecedented prospects of curing diseases caused...your research Browse plasmids for base editing References Anzalone AV, Randolph PB, Davis JR, Sousa AA,...
  19. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

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    ...nutrient-rich and also the best source for mycoplasma proliferation. Plus, it is added to the media after autoclaving... contact with a nutrient source, they start proliferating shortly. Once they are present in the laboratory...Bioluminescence assay kits are available from different biotech companies which are enzyme-based mycoplasma...and a Bachelor's degree in Biotechnology.       References Degeling MH, Maguire CA, Bovenberg MSS, Tannous...
  20. Finding Your Science Policy Path

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    ... away from the bench, I have worked for seven different organizations in a variety of science policy roles...cultural, and political. Science policy looks differently depending on how and where you pursue it. It ...you do science policy? Just like there are many different types of science policy careers, there are many...NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab a virtual...
Showing: 681 - 700 of 1064 results