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We narrowed to 968 results for: LAT;

Showing: 281 - 300 of 968 results

  1. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    ...continuing to copy the template rather than dissociating).  MarathonRT, isolated from Eubacterium rectale...fields of biology and enable the conversion of RNA templates to DNA but suffer from several important shortcomings...structured HCV genome (~9.6 kb of highly structured template!). MarathonRT also faithfully transcribes RNA ...protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find all of the nanobody toolkit ...biotin. The biotin-labeled proteins can then be isolated and analysed. In this work, the authors specifically...discovery of proteins involved in transcriptional regulation and chromatin structure at your locus of choice...technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements by restricted spatial tagging...

  2. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...with ssDNA repair templates. Their work demonstrates that different repair template strategies activate...using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template to increase ...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...working with CRISPR than just designing a repair template that contains your desired edit, and I want to...CRISPR updates on our blog to keep current with the latest and greatest! Addgene resources 3 Tips to Improve...Editing in Human Cells Hands-on tips for repair template design from guest bloggers Dominik Paquet and ...

  3. Mentoring for Scientists: I Have a Mentor, What Now?

    Type
    Blog Post
    ...mentor. How do you make the most of this new relationship? Consider adding formality and active goal setting... setting to your mentoring relationships, so that you can reap rewards in the form of reaching career ... career path is a great start to a mentoring relationship, but if you don’t take this past the chatting...meetings and setting the direction in a mentoring relationship falls on the shoulders of the Mentee. A Mentor...progress over the course of an ongoing mentoring relationship (in a book or online). Mentee: Make a list of...secret of accountability To make a mentoring relationship really productive, the most important step you...on setting SMART goals using a Development Plan template. A Development Plan or mentoring journal will ...

  4. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...efficiency of Cas9 cleavage is relatively high while the efficiency of HDR is relatively low, meaning that most...without double-strand breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime editing...summarize the major ones in Addgene’s repository later in the post. Advantages of prime editing Less constrained...homology-directed repair Homology-directed repair (HDR) stimulated by double strand breaks has been widely used ...increase activity, enhance binding between the template and primer binding site, increase processivity...remake that strand using the edited strand as the template. The PE3 system does just this by including an...PE = prime editor; RTT = reverse transcriptase template; PBS = primer binding site. Image from Doman et...

  5. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ...transduced by lentivirus and have been difficult to manipulate genetically. To solve this issue, Sarah Teichmann...disease is driven by atherogenic lipoproteins that accumulate within the vascular wall. ApoB, the main structural...lipoproteins, shuttles lipoproteins through the circulatory systems, but can also lead to the formation of... of ApoB-LPs has been shown to have a strong correlation with cardiovascular disease and indirect measurement...in vivo model to identify drug candidates that modulate ApoB-LP size and concentration, Steven Farber ...that the system is active in E. coli and closely related species, is amenable to use with other bEBP/UAS...and the system may not be adaptable to distantly related species from E. coli. This tool should serve as...

  6. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    ... AAV Targeting and manipulating specific, genetically-defined neuronal populations with intersectional...calcium sensors and GABA sensors) Calcium sensors The latest calcium sensors from Janelia (jGCaMP7) continue...potential, GCaMP7s is approximately twice as bright, relative to baseline fluorescence, as the GCaMP7f variant...useful for imaging signaling activity in large populations of densely-labeled neurons because background...all our biosensors inventory, and check out our latest vectors below: pGP-AAV-syn-jGCaMP7f-WPRE (104488...Optogenetics continues to be a useful method for manipulating specific neurons. We recently packaged a soma-targeted...specificity of influence measurements by reducing photostimulation of the axons and dendrites of non-targeted...

  7. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ... can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony ...thousands of times by propagating through an enormous lattice of identical subunits. A brief history of R bodies...coli in the 1980s  (Kanabrocki et al 1986). Much later, it was shown that putting the operon under a strong...mutants that couldn’t extend at a normal pH and later realized that these mutations clustered in a particular...their ability to penetrate other barriers (like vasculature or extracellular matrices) in response to pH ... response to environmental conditions. More speculatively, R bodies might endow divergent organisms with...Protein Piston That Breaks Membranes to Release Encapsulated Cargo. ACS Synth. Biol. (2016). doi:10.1021/...

  8. Exploring Applications of the Bioluminescent HiBiT Tag

    Type
    Blog Post
    ...proteins regulate vital cell and tissue processes, but their low abundance, complex regulation, and context-dependent...can include background autofluorescence from the plates or media, which can obscure weak signals; photobleaching...expression and localization. HiBiT-tagged targets in translational research Researchers can incorporate HiBiT with...difficult-to-study genes. Its main function is to regulate DNA repair, senescence, and apoptosis. Work by...at R273H to match a mutation frequently found in late-onset GC. HiBiT-TP53 expression was detected by ... of cystic fibrosis transmembrane conductance regulators (CFTR) (Kamada et al., 2023). CFTR is a membrane...., Robers, M. B., Benink, H. A., Eggers, C. T., Slater, M. R., Meisenheimer, P. L., Klaubert, D. H., Fan...

  9. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    ...gene a megabase away, or looking at the histone methylation status at a double-strand break that needs repairing...chromosomes interact with each other. Chromatin regulation is complex and influences--directly or indirectly...difficult to consolidate every conceivable chromatin-related plasmid in a collection that grows on a daily basis... Yi Zhang Leonard Zon  You!     Chromatin-related genes with plasmids available from Addgene The ...directly on our website. In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1...from Addgene Many articles reporting chromatin-related research describe the production of plasmids that...2018 Feb 13. pii: 1722491115. Addgene blog posts related to chromatin This is not the first Addgene blog...

  10. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    ... in an unregulated fashion and thus a marker for transfection. The other (mKate2) is regulated in the ...Constructs using the photodimerizer system CRY2-CIB1 to regulate botulinum neurotoxin were also created. The depositors...new light on mechanisms of miRNA-mediated gene regulation Article contributed by Michael Lemieux  Listen...microRNAs (miRNAs) are recognized as important regulators of post-transcriptional gene expression across...several years of research, the mechanisms of miRNA regulation have yet to be fully elucidated. Ron Weiss’s ... present, it will repress expression of mKate2 relative to EBFP2. Weiss’s group used these reporters to...define the different cells present in a mixed population, or identify differentiation and disease states...

  11. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...is software product management, and how does it relate to science? In the simplest of terms, a software...more about the stakeholders, the product managers collate and organize that information and form it into ...science backgrounds, most did not. It was my job to translate complex lab operations workflows into a format... help improve their work. Then, I distill and translate that valuable information back to the rest of ...science companies, try to find internships with translatable skills: working with data, working with people...jargon and know the concepts as soon as possible Later in your career:  Show demonstrated skill of people...project management  Bolster your skills in strong relationship-building and influence. You’ll need a large ...

  12. How to Make Friends and Meet People at a Scientific Conference

    Type
    Blog Post
    ...no better place for a scientist to make new relationships than at scientific conferences. Conferences ...this is a great ice breaker. Of course, real relationships go further and grow over time, but being interested...fun, memorable meal is a great way to build relationships. Make a list of experimental problems or questions...work – casual socializing is how people build relationships that are memorable. It isn’t always appropriate...overdo it (some of my best studies were designed over late night poker games at Gordon Conferences). You don... of connection. You can also see and offer congratulations on job changes and promotions.  Follow academic...scientists on ResearchGate and/or via Pubmed alerts. Congratulate your connections when they get a new paper published...

  13. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...ddifferent cell types, including difficult-to-manipulate primary cells, achieving knockdown within 1-2...methods for TRIM-Away delivery. For small cell populations, co-microinjection of TRIM21 protein and the ...the target antibody works well. In larger cell populations, you can deliver these components together using... electroporated cells re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines... CRISPR/Cas9-based system to study epigenetic modulation. While catalytically inactive cas9 (e.g. dCas9...-minute analysis of the kinetics of chromatin regulation and subsequent mathematical modeling. Its reversibility...amount of noise in reporter gene expression by modulating the induction of transcription factors 1 and ...

  14. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...or 16S rRNA isolated from the microbial community) reveals the RBS and allows translation of the reporter... Thus, cells can be both sorted using FACS and isolated with antibiotic selection. The team tested this...followed by a reporter gene. The hairpin blocks translation by sequestering the RBS, but binding of the “...relevant host biomarkers. The lab validated their platform on clinical stool samples by comparison to qPCR...Betaproteobacteria, and Gammaproteobacteria species isolated from different bee species. BTK can be used to...division in the gut microbiota Monitoring microbial population dynamics in the gut can provide insight into ... al. "A low-cost paper-based synthetic biology platform for analyzing gut microbiota and host biomarkers...

  15. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ..., and electroporated them with an ssODN repair template in induced pluripotent stem cells made from the...fertilization. Since Ma et al. designed a repair template with two synonymous mutations to distinguish it...were able to distinguish between repair from the template vs repair from the maternal MYBPC3 allele. Interestingly...from the single-stranded oligonucleotide (ssODN) template supplied by Ma et al., but instead from the mother...supplied single-stranded oligonucleotide (ssODN) template. Embryos also underwent editing through the NHEJ...NHEJ pathway. Created with BioRender.com. In later characterization studies, the edited embryos proceeded...embryos to undergo editing using the provided ssODN template. In fact, this finding led the authors to title...

  16. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ... in neurons reveals a mechanism for regulating its translatability." Science 343.6169 (2014): 419-422....Therefore, I set out to study how protein turnover is regulated in neurons. Protein turnover is the combined ...the abundance of proteins in cells. Homeostatic regulation of protein abundance is essential for normal ...acids. The use of radioactive materials is highly regulated, requires designated work areas and come with ...proper microscope setup we can observe the whole population of one particular protein in neurons or in any... has recently published techniques to follow translation in living single cells. However, before CRISPR-Cas9...synthesized proteins of interest were detected and later visualized by FUNCAT-PLA. Before these methods ...

  17. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ..., a researcher can cause them to interact by stimulation with blue light.  Similarly, light can also be... and is reversible upon termination of light stimulation. CRY2clust enables faster assembly and disassembly... and better distribution of clusters without accumulation in subcellular compartments such as nuclear ...Furthermore, CRY2clust, when integrated into a Ca2+ -modulating optogenetic system (OptoSTIM1), caused two-fold...Listen to the soCoChR podcast segment Optogenetic stimulation is a powerful tool used by neuroscientists to...artifactual PSCs commonly observed with direct stimulation are largely absent in the soCoChR-expressing ...rescued by the xanthine produced via complete demethylation of caffeine. For example, the Barrick lab was...

  18. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...Because of the availability of tools for genetic manipulations, mice have become the model of choice for many...experimenter blinding to avoid bias Genetic manipulation in behavioral neuroscience using viral vectors... is often done in conjunction with a genetic manipulation so that behavior can be linked to a specific... region. When it comes to making the genetic manipulation, there are two main decisions to make: 1. Which...inhibition upon light (or drug) application.  Genetic manipulation can also reduce expression levels of certain...spatial and temporal control of gene expression manipulation. They are injected using a stereotactic frame...depending on whether it is an overexpression gene, downregulation via RNAi or gene editing/deletion using CRISPR...

  19. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...and Sibernetic, a fluid mechanics simulator that can be used to simulate muscle tissue as it applies to ...of mRNA are important for post-transcriptional regulation in C. elegans. To study the role of 3’UTRs, the... C. elegans 3’UTRs structures, alternative polyadenylation for all protein coding mRNAs, and annotations...GAL-4 based bipartite expression system that can regulate gene expression in C.elegans. In this system a... to activation sequences upstream of genes to stimulate expression (Wang et al., 2016). In 2018, the lab...vectors that each contains a guide RNA and repair template of your choosing (Schwartz and Jorgensen, 2016...PMC3531189. Mangone, Marco, et al. "UTRome. org: a platform for 3′ UTR biology in C. elegans." Nucleic acids...

  20. Of Myc and Men

    Type
    Blog Post
    ...them in vivo and in vitro, especially in the mid-late 20th century with the advent of hybridoma technology...recombinantly append their proteins of interest with unrelated peptides that already had high quality antibodies...an amide group added to its C-terminus post-translationally, which turned out to be critical for antibody..., another group of researchers was working on isolating monoclonal antibodies to better probe the functions... c-Myc protein or the C-terminus, Evan, et al. isolated six different monoclonal antibodies (Evan, et ...sequenced the variable domains of Myc1-9E10 in the late 1990s (Fuchs, et al. 1997; Schiweck, et al. 1997...Evan GI, Lewis GK, Ramsay G, Bishop JM (1985) Isolation of monoclonal antibodies specific for human c-...
Showing: 281 - 300 of 968 results