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Showing: 41 - 80 of 1006 results

  1. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    ...article in the Addgene Blog Mentoring series will cover 2 of the 7 mentoring questions I set out to answer...programs A formal mentoring program can be a great way to get started with mentoring. Read this Catalyst...Catalyst Report “Making Mentoring Work” to learn about what makes a good mentoring program. If your organization...also a great place to practice your mentoring skills. Online mentoring can work if both parties are diligent...my upcoming mentoring post in which I describe how to organize a successful peer mentoring circle. Asking...School researcher Connie Cepko to learn about her mentoring style. Mentors at home and away We should all ...organization has a mentoring program, consider tapping into it and helping make it work by being an educated...

  3. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... genome engineering in bacteria, alongside more established tools such as Cas9-recombineering, CRISPRa...and bind target DNA sequences using RNA-DNA base pairing. Separately, TnsABC forms a transposase complex... two transposase proteins (TnsB and C). While requiring fewer components for RNA-guided DNA integration...).  CRISPR-transposons for bacterial genomes engineering In the Sternberg group’s initial report of VchINT.... For transposons to be specifically recognized during transposition, genetic cargos must be flanked by...currently studies CRISPR biology and CRISPR genome engineering technologies. Message or follow him on twitter...high-efficiency, multiplexed bacterial genome engineering. Nat Biotechnol. https://doi.org/10.1038/s41587...

  4. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    ...distribution is recovering from the pandemic-related slowdown. More scientists are sharing plasmids and ...are able... Thanks to everyone for being awesome during a tough time. We're all going to get through this...This is a time when we need as much open science sharing as we can get. The effects of Addgene’s brief...Addgene Team because they’re all doing amazing work during this difficult time. One function that rose to ...had to figure out a way to maintain our services during the crisis and maintain our workforce for when ...enough funds to make most of our payroll for the spring and early summer months. We are also extremely ...for grant-driven research being done in all labs sharing materials via Addgene. Perhaps if each of many ...

  5. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...without looking at what they’re delivering and where they’re delivering their payload to.   Figure...anniversary celebration, we’ve been finding and sharing stories unique to our repository and resources....prioritization happens for good reasons. We began offering our viral vector service because we realized that... are available quickly. Beyond the service This brings us to the final piece of the viral vectors service...our AAV Data Hub as a community platform for the sharing of that exact data. Here, requestors can upload...their experimental data in a standardized format, sharing how well a given viral vector worked — or didn’...save time in selecting experimental conditions. Sharing our experiences Addgene’s plasmid repository gave...

  6. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...roles. Therefore, institutions are responsible for ensuring that either all affected scientists are paid above...here) pushing for exemptions for postdocs, and lowering of the proposed salary level for exemption. Discussions... are institutions doing? Institutions are now preparing to comply with the changes to the law on Dec 1st...of 340 institutions with postdocs in science, engineering and health from the NSF’s 2014 Graduate Student...University of Delaware has a minimum of $36,360, requiring an increase of 30.6%. We have attempted to check...

  7. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ...removal helps prevent the invading genome from “recovering” its original sequence. If you’re looking to ... High Fidelty Cas9 Mutants Browse All Genome Engineering Posts Additional Resources on Addgene.org Check... Mary Gearing...

  8. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...base pairing, it has promise to be host-independent, beyond the oft published metabolic engineering realm...summarizes Zalatan JG, et al.'s recent paper, "Engineering Complex Synthetic Transcriptional Programs with...of research has shifted in recent years toward tinkering with regulatory networks and multiplex gene control... ‘biofactories’, most successes in metabolic engineering have been limited to the overexpression of various...scaffolds follows the rules of Watson-Crick base pairing, allowing for rational, straightforward design....scRNA design is readily apparent in metabolic engineering. The authors elegantly demonstrated the potential...violacein, a distinctive pigmented protein. By altering the transcriptional programs, the authors sequentially...

  9. Addgene @ Keystone: Thoughts on Precision Genome Engineering and Synbio

    Type
    Blog Post
    ...talk, which focused on the history of genome engineering from ZFNs through TALENs and CRISPR-Cas nucleases...huge impact on this field. Advances in genome engineering & CRISPR technology That afternoon's workshop...workshop, entitled "Advances in Genome Engineering", brought us 8 short talks on every conceivable way of examining...more parts-oriented synthetic biology. I've been answering biology customer service questions for nearly ...scientists are tackling this type of genetic engineering is a testament to how amazing the downstream ...downstream payoff could be. Though I attended an engineering school as an undergraduate, I never fooled myself ... The talks we heard Monday night represented engineering first and foremost, from the custom software ...

  10. Data Freedom: The Expansion of Data Sharing in Research Publications

    Type
    Blog Post
    ...jwoodgett.     More Discussions About Data Sharing: PLOS Data Sharing Policy Discussion PLOS’ New Data Policy...this year when it announced its data access and sharing policy. Since early March, the open access publisher...reasonable. PLOS has since clarified their data sharing policy and recently announced that of the 16,000...funding agencies (e.g. Wellcome Trust, NIH Data Sharing Policies, etc.) and is founded on the principle...data should be included? We should recognize that storing data in a sharable form is indeed a significant...Publishers have also contributed to this issue by requiring ever more condensed methods descriptions, legends...existing, published data. The importance of data sharing The reason this topic is important to my lab is...

  11. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    ... learning. 1. Create a peer mentoring group We don’t all get the mentoring support we need, and most of...Own Peer Mentoring Group: A How-To Guide for Scientists.   Once in-person, peer-mentoring groups ...use more mentors to listen, guide and support us during training and throughout our careers. An easy way...goals.  I’ve been surprised to see that virtual mentoring groups seem to be as successful as in person groups...? You could do a talk virtually for your peer mentoring group (see above). However, now that we are all...present, I’m asked what Addgene “looks for” when hiring a scientist. Of course, we have different criteria...

  12. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...Collection under the insect expression system or by filtering by fly as species of gene. Find Gal4 and UAS plasmids...select for transformed bacteria, but you might be wondering, “How do I know which of my flies have the desired...about Using Cre in Drosophila Read Other Genome Engineering Posts Use FLEx Vectors for Site-Specific Recombination... genome engineering...

  13. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...interference, these enzymes can knock down RNA without altering the genome, but Cas13s have higher on-target specificity...enzymes are the latest tools in the transcriptome engineering toolbox. How were Cas13d enzymes discovered?...MAPT, the gene encoding dementia-associated tau, delivering dCasRx and a 3-spacer array targeting the MAPT... tell how large the genome and transcriptome engineering toolbox will be. It is, however, certain that...that the impact of CRISPR scientific sharing will continue to grow, and we at Addgene appreciate our depositors...References Konermann, Silvana, et al. “Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors...  Mary Gearing...

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

    Type
    Blog Post
    ... developed during the COVID-19 pandemic. Our mission to accelerate research through sharing reagents has...2021.03.10.434840 Tsang J, LaManna CM (2020) Open Sharing During COVID-19: CRISPR-Based Detection Tools. The...detect SARS-CoV-2 nucleic acids. Mammalian targets During the 2003 SARS outbreak, Hyeryun Choe’s lab identified...-CoV-2 entry into human lung epithelial cells (Biering et al., 2021). In their search, they found mucins...inhibitors of SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV-2 viral...academic and industry scientists through plasmid sharing and more. As you’ve read above, some of these plasmids....     References and resources References Biering SB,  Sarnik SA, Wang E, Zengel JR, Sathyan V, Nguyenla...

  16. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...and its use as a genetic engineering tool is frequently called recombineering - short for homologous recombination-mediated...components of the Lambda Red recombineering system The lambda red recombineering system has three components...for generating dsDNA recombineering substrates to avoid errors introduced during PCR. ssDNA Substrate ...substrates can be generated by ordering synthetic oligonucleotides or denaturing short PCR products. Either..., Court DL. Recombineering: A Homologous Recombination-Based Method of Genetic Engineering. Nature protocols...Costantino, N. and Court, D. L. 2014. Recombineering: Genetic Engineering in Bacteria Using Homologous Recombination...method that can be used for cloning or genome engineering and is based on homologous recombination. It ...

  17. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...by Benoit Giquel. CRISPR has taken the genome engineering world by storm owing to its ease of use and utility...Waltz, 2016), the potential of CRISPR for genome engineering in plants is also being realized. There are a...approaches, these techniques are cumbersome, often requiring several rounds of selection to isolate plants ...plants with the phenotype of interest. Genome engineering, on the other hand, allows for targeted modification...will present a general overview of plant genome engineering using CRISPR, highlight the specific modifications...plants, and outline the various plant genome engineering tools that are available to academic researchers... Addgene. CRISPR components for plant genome engineering CRISPR can be used to knockout, activate or repress...

  18. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ...important role in scientific exchange. In addition to ensuring affordable access to materials, repositories provide...since 1996.       More Blog Posts about Scientific Sharing: The 10 Most Distributed Plasmid Technologies ...10 Years Data Freedom: The Expansion of Data Sharing in Research Publications ...

  19. Spurred by COVID-19, Addgene Bridges Plasmid Sharing Between Academia and Industry

    Type
    Blog Post
    ... to academic scientists. Expanding plasmid sharing during COVID-19 Realizing this limitation, we worked...members of the scientific community support open sharing. But, one critical part was missing when the COVID...setting.  In light of the need for basic research during the current pandemic, universities are showing ...cells. We teamed up with Ginkgo Bioworks to quickly bring these plasmids to the scientific community - both...available to industry scientists To make plasmid sharing more open for academic and industry scientists,...the past months, we’re excited to see our open sharing impact grow beyond COVID-19. But, we can’t do this...office to discuss the possibilities for expanded sharing. Is there a plasmid you're hoping will be come ...

  20. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...large scale cell mobility are essential processes during embryonic development. These are required for a...development of cell sorters together with the engineering of fluorescently labelled antibodies made it ...)? How do blood monocytes patrol blood vessels during immune responses (Auffray et al. 2007, Finsterbusch...relatively little tissue absorption, weak tissue scattering and small amounts of tissue auto-fluorescence...understanding the key roles of immune cells in triggering an immune response. New fluorescent proteins ...Dynamics of neutrophil migration in lymph nodes during infection." Immunity 29.3 (2008): 487-496. PubMed...Jackson G., et al. "Macrophage and T cell dynamics during the development and disintegration of mycobacterial...

  21. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    ...raise salaries prior to the injunction are still honoring these plans. We are now collecting data on the...who had a contract - negotiated by their union - securing higher salaries. This decision will carry over...geographical regions When we originally started gathering this information, many assumed that those living...Read Gary's Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating...

  22. Genome Engineering, CRISPR, and Research in Singapore: Interview with Wei Leong Chew

    Type
    Blog Post
    ...career path and why he chose to go into genome engineering. Chapter 1: Early research on CRISPR and what...with new project ideas, applying for grants, and mentoring. Chapter 5: Wei Leong Chew's lab management style... 6: Projects in Wei Leong Chew's lab. Genome engineering, gene delivery methods, AAVs, and more. Chapter... genome engineering...

  24. Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring

    Type
    Blog Post
    ...these extreme environments. Gearing up for sample collection at the hot springs My husband, Paul Blainey,...all of the remote areas, including one hot spring requiring an overnight hike in. Navigating challenges...the hot spring. Hiking all day and processing samples by night was fun but also very tiring. It feels...Lab at Stanford University became interested in exploring biological dark matter – large tracts of the microbial... these microbes. From 16S rRNA sequencing, hot springs were known to be diversity hotspots containing ...aquaticus bacterium – was discovered at Octopus Hot Spring at YNP, and since then, many researchers have searched...temperatures and unstable ground around these hot springs make them extremely dangerous, we used telescoping...

  25. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ...Addgene Blog Mentoring for Scientists Series. I have been thinking a lot about Mentoring for over 10 years...group mentoring format could work wonders.  I have now seen the formation of hundreds of mentoring groups...also serve as a great first-time mentoring experience. They make mentoring less mysterious and train the ...useful for future mentoring relationships.   If you would like to start a group mentoring program at your...interested peers and form your own peer mentoring circle. In a Peer Mentoring Group all participants act as both...and what you want to cover during the year. Suggest that each person bring 3 topics and write them on ...involved in Peer Mentoring relationships? Comment below with ideas from your own mentoring group or with ...

  26. "What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring

    Type
    Blog Post
    ...to learn about her mentoring style. Top 7 questions scientists ask me about mentoring In this blog series...the other six mentoring questions in future posts. If you have a question about mentoring that you want... enough?  For many years I’ve been organizing mentoring programs for scientists and doing training to ...fulfilling, productive relationships. I will be sharing what I’ve learned along the way in this ongoing... to the top questions scientists ask me about mentoring. Here they are: What makes a good Mentor? How...? What is this I hear about “group” or “peer” mentoring? How do I ask someone to be my Mentor? I have ...certainly realized the importance of seeking out mentoring as an important aspect to their ongoing success...

  27. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...bacterial CRISPR engineering One common way of accomplishing bacterial genome engineering is done with recombineering...combines CRISPR and recombineering-based MAGE (Multiplex Automated Genome Engineering) technology (Ronda...combines recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang...promoters in Pseudomonas syringae and Klebsiella oxytoca. Microbiome engineering Scientists have begun to...discovered in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria...quirks change the way CRISPR-mediated genome engineering functions in bacteria, but have no fear - plasmids... recombineering, a technique that utilizes phage recombination machinery to promote homologous recombination...

  28. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    ...KTRs are not well suited for monitoring kinase activity that occurs during nuclear envelope breakdown (...important for regulating signal transduction and measuring kinase activity at the single-cell level can aid...signaling activity and cell phenotype. One method for monitoring live single-cell kinase activity is FRET, but...Advantages of KTRs When compared to FRET reporters for measuring kinase activity, KTRs have two key advantages....

  29. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

    Type
    Blog Post
    ...multiplex genome engineering. Image from Sakuma at al. (2015). Multiplex genome engineering demonstrated...system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to researchers at Japan...making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that gap. As Sakuma and...efficient viral vector-mediated multiplex genome engineering. The system now provides an efficient targeting...Sakuma’s protocol. Reference: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector... multiplex genome engineering...

  30. Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

    Type
    Blog Post
    ...database and ordering platform soon to be released. For individuals interested in ordering from ABRC for...Learn how BioBuilder can bring Synthetic Biology into the classroom Engineering the Plant Genome Using ...to us at [email protected] for help setting up an ordering account. Using Arabidopsis in the classroom has...CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org Find Plant Vectors...

  31. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...platform for sharing, annotating, and storing life science protocols. Since we began sharing this protocol...Developing Lab Management Software    Organizing and sharing your updated protocols So that’s great, but what...Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check out all the benefits...feature further simplifies lab work if you like to bring your phone in the lab, though, honestly, this still...non-bench work – but is there a sterile, clean way to bring your phone into the lab? Maybe having a dedicated... protocols listed on OWW, including those for preparing your own competent cells. If you’re familiar with...exchange currently boasts over 2,500 protocols covering a variety of bio and biomedical topics. Protocol...

  32. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...using glass cloning rings to isolate colonies of antibiotic-resistant cells; during the time required to... to grease, place and fill a dozen cloning rings, the remainder of the colonies on the plate dried out...out and died. The alternative to cloning rings, dilution cloning into 96-well plates, is extremely time...minutes- a far more rapid pace than using cloning rings. A picture of a 60 mm dish ready for picking is ...differ grossly from those isolated using cloning rings in terms of growth rates, morphology or other physiological...

  33. CRISPR Guide

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    Collection
    ...scientists have modified for a wide range of genome engineering techniques . Collectively, these techniques are...R epeat) technologies. Before CRISPR, genome engineering approaches like zinc finger nucleases (ZFNs) ...CRISPR rapidly became the most popular genome engineering approach. Figure 1: Overview of the basic CRISPR...first and come back to HDR . NHEJ is a naturally occurring repair pathway and is the primary repair mechanism... for DSBs. While this pathway is efficient in repairing DSB, it frequently causes small nucleotide insertions...: Double-Strand Break (Cut) Multiplex Genome Engineering Many CRISPR experiments involve editing more ...more than one gene. In these cases, delivering multiple gRNAs using a single plasmid ensures that all gRNAs...

  34. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ... O 2 ) Monitoring of basal H 2 O 2 levels with peroxiredoxin-based probes Real-time monitoring of basal...81. Loren Looger Calcium NEMO calcium sensors Engineering of NEMO as calcium indicators with large dynamics...sensors, ER-targeted CEPIA1er ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in mammalian...sensors ZapCY1 and ZapCY2, Golgi or ER targeted Measuring steady-state and dynamic endoplasmic reticulum...alternately colored FRET sensors for simultaneous monitoring of Zn(2)(+) in multiple cellular locations. PLoS...Improved ATP FRET Sensor For Yeast Shows Heterogeneity During Nutrient Transitions. ACS Sens. 2020 Mar 27;5(3...-0. Yulong Li ATP/ADP Monitor the ATP:ADP ratio during live-cell imaging (Perceval) A genetically encoded...

  35. AAV Packaged on Request

    Type
    Collection
    ... high-quality quality control measures such as titering, purity assessment, sterility testing, and viral...Request AAV Packaged on Request Overview Details Ordering Share Your Results Eligible Plasmids Additional...levels. We offer 963 in-stock viral vector preps, delivering high quality and ready availability at affordable...Addgene’s comprehensive quality control process: Titering Purity assessment Sterility testing Viral genome...chosen plasmid on its material page. Request and Ordering Process AAV production will take approximately...requests will be approved within two business days! Ordering At your convenience After approval, the next step...AAV Production 6–8 weeks Quality is our priority. During this time, we amplify DNA, produce the prep, and...

  36. Viral Production

    Type
    Collection
    ...customers. Details about our production protocols, titering methods, and quality control are described below... or through collaboration with viral vector manufacturing facilities, such as the University of Pennsylvania...are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or by the University...University of Pennsylvania Vector Core. In general, titering is performed by the facility that produced the...those achieved using the standard qPCR method. Titering by the University of Pennsylvania Vector Core ...Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral...aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral preparations that have...

  37. Zinc Finger Consortium Reagents

    Type
    Collection
    ...nuclease fusions Related Genome Engineering Technologies TALEN Engineering Reagents CRISPR Plasmids...Find reagents for engineering and expressing zinc finger proteins deposited by Zinc Finger Consortium...have deposited at Addgene various reagents for engineering and expressing zinc finger proteins for distribution...reagents are provided below: Zinc Finger Genome Engineering Reagents Zinc Finger Arrays for Zebrafish Genes...fingers targeting specific genes Oligomerized Pool ENgineering (OPEN) Reagents - Engineer zinc finger arrays...arrays via OPEN method Modular Assembly Engineering Reagents - Engineer zinc finger arrays via Modular Assembly...

  38. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    ...bad news. The bad news is that some of tubes’ o-rings are, in fact, permeable to CO2 at low temperatures...What we found Not all tubes (or rather, not all O-rings) were permeable to CO2 at low temperatures. We tested...Conclusions and handling recommendations Not all O-rings function optimally at -80°C. For samples that are...or not the dry ice we use to keep viruses frozen during shipment was having an impact on the samples. We... Order Ready to Use Viral Preps from Addgene Measuring the pH of our samples: What we did To measure ...CRIPSR User (By a 1st Time CRISPR User) Tips for Titering Your Lentivirus Preps Additional Resources on...

  39. Antibody Production

    Type
    Collection
    ...thaw of antibodies and suggests preparing single-use aliquots before storing at -20 °C. Each antibody catalog...production, all antibody preps are titered and undergo stringent quality control by Addgene or a trusted quality...distribution. Details about our production protocols, titering methods, and quality control are described below...Coomassie staining, the sample is separated via a denaturing SDS-PAGE gel alongside serial dilutions of a ...each order based on the total items in the order, ensuring each item ships safely and most economically. ...

  40. Plasmid Collections

    Type
    Collection
    ...before starting your experiment. Genome Engineering Genome Engineering Overview CRISPR Cre-lox TALENs Zinc...Kit Types Fluorescent Proteins Cloning Genome Engineering Pathways & ORFs Synthetic Biology Pooled Libraries...plasmids all built with the same backbone and only differing in a small region. Click here to view all pooled... Cell Migration Consortium Center for Genome Engineering Impact of Genomic Variation on Function (IGVF...
Showing: 41 - 80 of 1006 results