We narrowed to 1,018 results for: LEA
-
TypeBlog Post...DNA could lead to transgene expression in recombinase-negative animals, producing misleading results. ...the plasmid used to create the viral prep. In a clean viral prep, like AAV-44362 shown below, more than...from these known impurities, as was the case for clean sample AAV-44362. In addition to these expected ...number of hits to the same sequence (gene). In a clean sample, there are typically fewer than 10 hits to...receive the best AAV vectors for your research. Please use the comments section below to let us know if...Topic Page Subscribe to Viral Vectors Blog Posts Learn How We Use NGS for Plasmid Quality Control Resources...
-
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post...that delivering RNA-guided nucleases or TALENs together with AdV donor DNA leads to a vast majority of AdV-modified...chromosomal region. The CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various... and demonstrated by Manuel Gonçalves and his colleagues at Leiden University Medical Center are now available...DNA for accurate genome editing with engineered nucleases. Nat Methods 11:1051–1057 . https://doi.org/10.1038...Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse...delivery of CRISPR/Cas9. Journal of Controlled Release 298:128–141 . https://doi.org/10.1016/j.jconrel... -
PITChing MMEJ as an Alternative Route for Gene Editing
TypeBlog Post... PITCh plasmid to release the cassette, and an insertion-locus-specific gRNA cleaves genomic DNA. These...it’s become clear that the power of this editing platform lies in the diversity of nucleases and their .... For more mechanistic details on HDR and NHEJ, please see the linked blog posts. Compared to its counterparts...compare to CRISPR sticky-end insertion using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, ...Experiment Additional Resources on Addgene.org Learn All about CRISPR on Our CRISPR Guide Pages Find ... -
Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments
TypeBlog Post...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...this plasmid, and if it still being used. After learning of it's continued popularity, I decided this protocol...thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem many scientists... found in our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check...lab for non-bench work – but is there a sterile, clean way to bring your phone into the lab? Maybe having...protocols to the site and will keep you updated as I learn to use the resource. Nature Protocol Exchange Website...I missed, let me know in the comments below and leave your thoughts on how the app works for your lab’... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post... Developed by David Liu and colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically... to this system by engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing...control for your experiments. For more information, please see Addgene’s information page for the Rinehart...neuronal signaling. For more information on DREADDs, please see the Roth lab's DREADD users blog. New ... expression plasmid that was used by Chen and colleagues for genome editing inXenopus tropicalis. (Guo...information on using the Platinum Gate TALEN Kit, please see the Yamamoto lab’s protocol for TALEN construction...mGRASP A Nature 2012 article by Jinny Kim and colleagues describes their efforts to map the location and... -
CRISPR 101: Cytosine Transversion Editors
TypeBlog Post...editors. They adjust the side groups of DNA bases and leave the core ring structure intact, allowing them to...guide RNA to base pair with the DNA target strand, leaving the non-target strand unpaired and accessible on...it the “edited strand.” Uracil excision repair leads to the first base transversion editor The story ...improved the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked...included in the CGBE. They also developed a machine learning model capable of predicting editing outcomes of...that were already accessible. It's not entirely clear why this is, but it might point to a more dynamic... screens, target-library analysis, and machine learning. Nature Biotechnology, 39(11), 1414–1425. https... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog Post...to see the dirty laundry exposed here, but it’s unclear how much they will actually care. The section of...technology. Though these case studies are “old,” they lead Zimmer to one of his most critical observations:...detection had over 1,000 authors. The chapter on deep learning addresses the dilemma of creating a technology...discussed The State of Science with the five Addgene colleagues who read it with me, the consistent response ...non-scientist alike was that the book left them wanting to learn more. There are individual paragraphs that may have...task to set oneself. As a scientist who enjoys learning a little bit about many different things (part...about a long time. For the scientists who like to learn everything about one specific topic, The State of... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog Post...several washes, this tag can be cleaved by the bdNEDP1 protease, thus releasing purified nanobodies from the...common protein purification methods. Read on to learn more about nanobodies and how their structure and...primary antibodies raised in different species, or at least from different IgG subclasses, so that secondary... and the target antigen to about 2 nm, which can lead to higher resolution for super-resolution microscopy...rabbit nanobodies. If you’re interested, you can learn more about how nanobodies are identified in this...University Use Nanobodies to Control Protein Activity Learn more about protein tags Read about E. coli strains... -
Lentiviral Vector Uses and Overview
TypeBlog Post... clinical gene therapy applications. Read on to learn more about the current (and future) applications... co-transfection of these plasmids, 293T cells released transgene-containing lentiviral particles into...and they showed that expression persisted for at least thirty days. Although adeno-associated viral vectors...activation from the 5’ LTR) and Rev (facilitates nuclear export of transcripts). A different viral envelope...safer, this system may be more cumbersome to use and lead to lower viral titers due to higher plasmid number...Addgene Blog Your Lentiviral Plasmid FAQs Answered Learn How Lentiviral CRISPR Libraries Enable Genome-Scale... on Addgene.org Check out Our Lentivirus Guide Learn about Virus Biosafety Find Lentiviral Vectors for... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog Post...journals. In addition, it is my pleasure to interact with my colleagues in meetings of scientific societies..."Biology is Method", i.e. new technologies often lead to big discoveries in biology. Therefore, my lab...translocation trap (ITT) system to analyze signal-induced nuclear translocation of proteins in 2004. In addition,...much enjoyed interactions with many excellent colleagues at NYU and visiting scientists, such as seminar..., I've been very lucky to work with excellent colleagues in my lab. On the other hand, I feel that researchers... conventional research. So, I hope my younger colleagues may be able to develop new technologies, hopefully... -
Career Insights: Technical Support Specialist
TypeBlog Post...the progress of the project. After that rather unpleasant experience I thought: I could do this job so ...and disadvantages of this overall rewarding role. Learn about Other Addgenies and Their Careers in Our Career...Technical Correspondent or similar. While there isn’t a clear distinction between these job titles, they may be...particularly tricky cases, team discussions can often lead to otherwise unobtainable solutions. To some extent...support can add a variety of skills to your CV and lead to other job opportunities (though it is not uncommon...Occasionally it can also be necessary to take the lead and be assertive. You develop an understanding of... Now? Academic v. Non-Academic Career Decisions Learn about Science Career Options Explore Tips for Technical... -
How to Write a Scientific Review Article
TypeBlog Post...define your scope from the onset. Perhaps you, a colleague, or your adviser was invited to write on a particular...to get a sense of the themes in the field and to learn what people cared about. I also used reviews to ...beginning I felt like this was cheating. Eventually I learned that it’s practical because there simply isn’t ...enough time to read every single paper. I also learned to appreciate the discussion sections of the primary... down and get anything done, so unless I had at least a two hour block, I wouldn’t work on the review....free two hour block of time was. Eventually, I learned that I was most productive in short bursts (even... could find the original paper if you wanted to learn more. Another example: we made a schematic of all... -
5 Reasons to Use Reddit for Science Communication
TypeBlog Post... science trainees still have to lead on their own development. Please see the AMA itself for all sorts... a great way to establish yourself as a thought leader and have real conversations with other scientists...forums and try to keep discussions on-topic and, at least sometimes, civil. r/Science in particular hosts ...answered many questions in the AMA but, as you’ll learn below, even with many resources at her disposal,...potentially hostile posters (this advice included leaving out the term “implicit bias” which is apparently...academic one for the majority of scientists and I am pleased to see graduate and postdoc programs beginning ... -
Tips for CRISPR Gene Editing in Mice
TypeBlog Post...target sequence was digested by sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination...there has less chance of contamination with RNase leading to degradation that can set you back days. Working...(e.g. face mask, and a dust free and specially cleaned work bench). Once the pups are born It takes roughly... variety of phenomena, it’s possible that we’ll learn much more from our favourite mammalian model organisms...off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1 (2014): ...Daisuke, et al. "Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9...Engineering with CRISPR Validate Your Genome Edit Learn about Non Homologous End Joining and Homology Directed... -
Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs
TypeBlog Post...nonimmigrants). The foreign worker must possess at least a bachelor's degree or its equivalent and state ...books, journals online and/or in print. Having a lead role or critical role at a prestigious institution...successes in the performing arts- not relevant at all. Please do not include the fact that you are a classically...company you must show that your company employs at least 3 full-time people engaged in researcher activities...from the point of view of experts in your field. Please remember, this is not a job reference letter. This...awards, presentations, in .pdf format. Last but not least, NETWORK! Networking is extremely important for ...business development, finance, and operations. To learn more, visit www.propelcareers.com. Casseus Law: ... -
The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs
TypeBlog Post...perhaps there is a clear winner, is there something that we can invest our time into learning and extract the...timers: You need to learn the language and what the parameters represent. The best learn-by-doing tool is...for improvement and a year later it became quite clear that others thought the same. Since January 2013...microhomology aware”. Trends: Fewer tools are being released with advanced functionality, it seems these kinds...blogger Cameron MacPherson! Cameron R. MacPherson is Lead Data Scientist for the Milieu Intérieur project,..."DNA targeting specificity of RNA-guided Cas9 nucleases." Nature biotechnology 31.9 (2013): 827-832. PubMed... -
Bringing Sustainable Practices to the Lab: Easy Action Items
TypeBlog Post...You wouldn’t leave your oven on all day to save time making dinner at night, so why leave your lab equipment...Addgene blog Learn about recycling initiatives for the lab Subscribe to the Addgene blog Learn about other...difference? Individual changes to our behavior can lead to cultural shifts that can have bigger impacts....even one less run a week makes a big difference. Clean out freezers so you can make room and don’t have... -
MassBioEd Career Exploration Day at Addgene
TypeBlog Post...High School visited our labs in Cambridge, MA to learn about the wide variety of science careers that Addgenies...that it’s never too early to start networking! Learning about careers at Addgene Afterwards, the students...: “Kory was awesome!”. The students also got to learn about intellectual property from our Tech Transfer...career paths, Michelle Healy said “You are always learning and that means it is possible to switch your field...Additional Resources Visit the MassBioEd Website Learn more about Dennis-Yarmouth Regional High School ... Blog Posts Get Career Advice from Our Bloggers Learn How to Improve Your Networking Skills ... -
Transferable Skills Guide: Time Management
TypeBlog Post...this gap by both highlighting that researchers do learn MANY transferable skills while working in the lab...have the best time management skills, but I’ve learned a lot since my days as an undergrad working with... variety of projects. I was therefore forced to learn a variety of ways to manage my time. As a busy researcher...distractions: Probably the easiest way to do this is to leave your phone in your bag on “Do Not Disturb” during... recommend the book, “Bored and Brilliant” for learning more fantastic reasons to remove yourself from...you have any other time management suggestions, please be sure to note them in the comments! Additional...Get Advice on Building a Grad Student Community Learn about Careers in Science Communication Resources... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog Post...fluorescence. In Brainbow 2.1, GFP is tagged with a nuclear localization sequence, allowing it to localize ...mouse generates cells that randomly express either nuclear GFP, membrane CFP, cytoplasmic YFP, or cytoplasmic...Cre undergoes exactly one recombination event, leading to each copy of the transgene expressing either... tags instead of XFP to optimize it for tissue clearing, a method that makes tissues, such as the brain...visible. Chemical tags are more resistant to tissue clearing methods and can be labeled by synthetic fluorophores...blog Read an introduction to live cell imaging Learn more about the origin of the Brainbow system Visit...Browse Addgene’s fluorescent protein collection Learn about fluorescent tools for in vivo imaging ...