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We narrowed to 1,000 results for: Eras

Showing: 301 - 320 of 1000 results

  1. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ...more hands on While teaching courses about gene therapy and genetic manipulation, I and other professors...absorbed if they don’t get a chance apply it in an interactive setting. The problem is…how can you make a “...similar types of thinking go into developing gene therapy strategies for various diseases. That’s when I ...-on”, but as much as I try being compelling, interactive, and application-driven to boost curiosity, I...skills to apply those concepts and understand how therapeutic strategies work. Find Addgene Blog posts about...pick the best strategy and propose a final gene therapy approach for the disease to be openly discussed... this way, the students also had to practice transferable skills like team building, collaborative critical...

  2. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...gene therapy have also been in the limelight. The FDA approved the first oncolytic viral therapy in 2015... science research all the way to clinical gene therapy. But how can you use these amazing tools in your... how we understand what genes do! Viruses have several options to allow you to do exactly that: shRNAs...integration of a plasmid. Viral transduction is also generally less toxic to cells than transfections, preserving...libraries makes them the ideal vector for the job. Therapeutic applications Vaccine development Attenuated and...nucleic acid-based vaccines. These vectors provide several advantages over traditional vaccines, including...being exploited as delivery mechanisms for gene therapy. The tractability of AAV, low immunogenicity, and...

  3. March for Science

    Type
    Blog Post
    ...17). Federal funding for science has been decreasing for the last five years and the first federal budget...passion for photosynthetic communities, microbial interactions, and science education.  Disclaimer: The views... in education, government, and civilization in general (1). Science and politics? Science is an apolitical... human consumption in comparison to 37% of the general public (5, 6). Similarly 90% of active research...mostly due to human activity while much less of the general public agrees (5, 6). Many of the new administration...only be by advocating for science education and literacy that this changes.  2) Lumping all scientists ...marching to advocate for science education and literacy as I believe people that can see and understand...

  4. Gendered Innovations: Why Does Sex of the Cell Matter?

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    Blog Post
    ...recipient interaction should be tested before being ruled out; there are many factors that interact with sex...strategic approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on...wrong costs lives and money. According to the US General Accounting Office, between 1997 and 2000, ten drugs...analysis of the US Women’s Health Initiative Hormone Therapy Trial, for example, found that the study saved ...Reduced use of estrogen plus progestin hormone therapy resulted in 76,000 fewer cases of cardiovascular... find 263,000 more osteoporotic fractures. The overall economic savings to the US healthcare system was... cells and tissues, specifically in stem cell therapies. Let’s go back to why ten drugs were withdrawn...

  5. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    Blog Post
    ...year and how these plasmids have been used to accelerate COVID-19 research in other labs. Spike protein... protein has had many opportunities to evolve. Several variants are now circulating and plasmids containing...from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding different spike...used these pseudoviruses to determine how effective sera from vaccinated individuals are at neutralizing ... the reverse transcriptase, protease, and Bst polymerase constructs from their assay to Addgene.  Drew...expressing MMLV reverse transcriptase and Taq polymerase that can be used in a one-step RT-qPCR reaction...SARS-CoV-2 researchers to investigate how the virus interacts with human cells. Addgene’s mammalian target plasmids...

  6. Hot Plasmids - October 2020

    Type
    Blog Post
    ...researchers tested several scenarios from embryo development to myeloid proliferation in acute myeloid ... IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation and had ...particularly challenging as they are great in number, generally very small, and can contain different neurotransmitters...proteins coupled with Fibronectin intrabodies generated with mRNA display (FingRs), Xue Han’s lab has ...cell cycle attempt to understand development, regeneration, and certain disease states. However, it’s difficult...proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in media without IL-3...withdrawn became entirely red as they could not proliferate. Eastman et al. Cell Reports 2020. https://doi.org...

  7. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase increases levels...transcription, also includes light-regulated histone methyltransferases and deacetylases. CRISPR and epigenetics ...initiates cytosine demethylation of DNA. However, several proteins in the DNA oxidation and repair pathways...delivery systems. Transcriptional repression DNA Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited...repression. Created with BioRender.com. DNA Methyltransferase MQ1 Margaret Goodell's lab has deposited pcDNA3.1...Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase from the prokaryote Mollicutes spiroplasma...

  8. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...There are several facilities nationwide where you can obtain excellent quality, high titer rAAV (including...including Addgene!), but you can also generate rAAVs in your own lab with standard molecular biology tools...our ddPCR for AAV quantitation blog post.  We generally do not recommend using ELISA-based titering assays...inverted terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org/10.1089... of gene expression in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https://doi.org/10.1186...can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038/sj.gt...Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. https://doi.org/10.1089...

  9. MXS Chaining

    Type
    Blog Post
    ...the functions of different genetic elements to generate non-natural tools such as novel biological circuits...and more. By combining these components, one can generate constructs to visualize cell cycle dynamics, titrate...applications. In one example, modular constructs generated with MXS-chaining were used to label subcellular..., MXS Chaining gave researchers the ability to generate the four initial unique constructs and later combine...ligation, the original pattern of the MCS is regenerated at the flanks of each ligation product, which...that the restriction sites of the MCS are only regenerated at the 5’ and 3’ termini of the ligation product...added to the 5’ or 3’ terminus using the newly generated MCS. MXS-chaining also has the advantage of being...

  10. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    ...providing many of the details not generally captured in the literature, scientists can use the Data Hub...commonly encountered in human gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org...Working with AAV vectors in the brain and peripheral nervous system presents a special challenge to scientists... process is often not detailed in the primary literature, and as a result can leave scientists under-informed...time and effort to test and optimize our vector - overall, I spent two years testing and reformulating our...dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https://doi.org/10.1038/s41434... Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https://doi.org/10.1089/hum.2009.169...

  11. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...FLEx cassette using next-generation sequencing (NGS) (Guerin, K. 2020). In general, we observe that 0.1 -... in the Central and Peripheral Nervous System and Relevance to Human Gene Therapy. Front Mol Neurosci ...FLEx Vectors Important Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting...Vectors). While this strategy greatly reduced the overall level of off-target transgene expression, low levels...greater off-target expression just due to more overall viral particles getting into those cells. Therefore...transduces your cells of interest. We recommend testing several different serotypes to determine which one is the...course experiment to check expression levels over several weeks allows the researcher to determine when the...

  12. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...begin to use CRISPR as a therapy. Low frequency off-target effects may generate irreproducible data in ... CRISPR experiments, many are turning to next-generation sequencing options over older assays. Biased... search criteria are not exhaustive. Mismatch tolerance settings are often limited to off-target sites...sites of <4 bp. The off-target list is also generally weighted by the position of the mismatch along the ...CRISPR editing in the model cell line or organism. Several options exist for unbiased off-target detection...labeling, enrichment on streptavidin and next-generation sequencing (BLESS) (Crosetto et al. 2013). GUIDE-seq...consistent set of standards as the field seeks to generate reproducible, quality data on the role of genetic...

  13. Plasmids 101: Origin of Replication

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    Blog Post
    ...plasmids. Growth conditions The amount of aeration, temperature, culture volume, antibiotic, and medium ...survive within cells. The replicons of plasmids are generally different from the those used to replicate the... to make additional copies. ori sequences are generally high in As and Ts. Why, you ask? Well, A-T base...pairs can be separated more readily at lower temperatures and allows the replication machinery room to...few copies depending on how they are regulated. Generally, control of replication is referred to as "relaxed...exclusively in E. coli? Gram negative bacteria in general? Both Gram negatives and Gram positives? Will you...compatibility with one or more other plasmids. Generally speaking, plasmids with the same ori are incompatible...

  14. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...structural studies of E3 ligase cereblon By Mike Lacy Several types of drugs target the ubiquitin E3 ligase cereblon...Lon and TBD domains, part of the HB domain, and several mutations to improve solubility and stability (...and mScarlet-I3, Oliver Hobert’s lab recently generated new constructs specifically for C. elegans (Cao...showed greater photostability and also matured considerably faster, with expression levels comparable to...notorious for its high resistance to antibiotics. Several CRISPR/Cas-based systems have been developed for...mechanosensor called PILATeS, Pulling-Induced Luciferase Association Tension Sensor (Zhong et al., 2024...sensor STReTCh, the 11-amino acid BiT fragment of luciferase is embedded in the I10 domain of the human titin...

  15. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...microglia The REVeRT Dual AAV Vector System Neurodegeneration Collection updates   Multiplexed perturbation...2024 under a CC-BY-NC-ND license. In this iteration, the size of the lentiviral insert was minimized...ensure this tRNA processing only occurs on the RNA polymerase III transcript, the sgRNAs were encoded on the...recombinant versions of mouse monoclonal antibodies generated and characterized by the Geiss Lab in their effort...A122D mutation in the rhodopsin domain conferred several new improvements. The new variant has increased...valuable tool for microglia research and broader therapeutic applications.  Find microglia-targeting AAV plasmids...10.1101/2023.12.12.571321.  Revolutionizing Gene Therapy: The REVeRT Dual AAV Vector System By Vaibhav Kawde...

  16. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ... We’ve previously discussed some of the general considerations for performing behavioral tests, but now... modulating. Viral vectors, in contrast, are considerably more versatile and specific in how they manipulate...field box or a circular corridor apparatus using a camera or tracking software. Locomotion is a highly complex... of individual neurons and in terms of their interactions with one another. In addition to the neuronal...controlling muscle activity and coordinating movement, several additional brain areas that reflect motivational...sensitization and might open new ways for a novel therapeutic approach against drug addiction. Anxiety tests...rodents can be used to study anxiety levels in general, or anxiety associated with psychiatric diseases...

  17. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...One of the biggest barriers to gene therapy treatment of central nervous system (CNS) diseases is the...because in vivo transduction of AAV serotypes generally tracks between strains of mice. This suggests ... engineered PHP.B. PHP.B was created by first generating a diverse library of AAV9 mutants that had a ...inheritance pattern of PHP.B permissiveness. The first generation of outbred mice (F1) had half the level of CNS...transduction as their C57BL/6J parent. The second generation (F2) of outbred mice also showed a Mendelian ...galactose and AAVR to transduce cells. The Wilson Lab generated results similar to the Deverman Lab when they ...Transport across the Blood-Brain Barrier." Molecular Therapy 27.5 (2019): 912-921. PubMed PMID: 30819613. Hordeaux...

  18. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...that stably express a gene of interest. While the general approach for using lentivirus, infect and select...consider is the age of your culture post thaw. In general, the higher the passage number of a culture the...routinely test for mycoplasma contamination. There are several commercially available mycoplasma detection kits...parameters. For example, while freeze thaw is generally not recommended, Holic et al. found that lentivirus...your target cells Labs tend to follow the same general procedure for lentiviral infections, produce the...earliest steps in lentiviral transduction is the interaction between the viral envelop and the receptors on... on the target cell surface. The possible interactions and hence possible targets for a lentivirus are...

  19. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...enhancers, and cross-cassette interactions in gene expression, and to generate multi-gene knockouts. Mammalian...MoClo background MoClo is a clever way to assemble several DNA parts in a specific orientation using Type ...2011). It can be performed at multiple levels to generate anything from individual transcription units (...products without the undesirable side products generated during chemical synthesis of these molecules. ...bacteria. If your yeast experimental goals are more general in nature, or you aren’t planning on using K. phaffii...Multiplex Yeast Toolkit builds on the YTK and adds several additional capabilities, including conditional ...., 2014). This kit is a great starter for your general plant cloning experiments. Addgene also has two...

  20. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...photobleaching. Interaction with receptors: Understanding how fluorescent dyes interact with specific receptors...intermolecular FRET for ligand-receptor interactions, require specific considerations regarding the choice of fluorescent...crucial in the world of biological research, medical therapy, and even drug development (Grimm & Lavis, 2022...targets through fluorescent labeling. There are several advantages to using these self-labeling proteins...is bound through green light emission. Key considerations in selecting fluorescent dyes/ligands Effects...best practice to determine optimal DOL is through several small-batch experimental labeling. Size is also...under certain conditions, which is an important consideration for assay design (Breen et al., 2016). When ...
Showing: 301 - 320 of 1000 results