We narrowed to 849 results for: NTS
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TypeBlog PostPublishedAug. 4, 2020, 1:15 p.m....the pandemic, it’s hard to get these reagents. However, when reagents have been made available through a...because they provide rapid access to validated reagents, especially during a time when these materials...Everyone. Case study 1: Cell lines Many of the reagents used to investigate SARS-CoV-2 were developed ...endeavor. Case study 2: Plasmids Another example of reagents that have broad applications but have proven useful...plasmids and resources! Case study 3: Mouse strains Reagents developed during the SARS-CoV pandemic more than...from the scientific community. Deposits of new reagents, policies from journals and granting agencies ...for storing, validating, and distributing such reagents are all ways in which the scientific community...
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Viral Vectors 101: Inverted Terminal Repeats
TypeBlog PostPublishedOct. 19, 2023, 1:15 p.m.... model. AAV plasmids contain several important elements, but among the most important are the inverted...inverted terminal repeats (ITRs). ITRs are the elements that make an AAV plasmid an AAV plasmid. Within the... and initiate replication. ITRs are cis-acting elements, which means that they are in the same DNA molecule...types Since ITRs are one of the most important elements of AAV and rAAV vectors, modifications can drastically...prevent unwanted mutations? During propagation, mutants rapidly replicate in a growth culture and result... sequencing is also helpful in quantifying ITR mutants since analysis of individual reads allows for identification...the Addgene blog Viral Vectors 101: Viral Vector Elements Viral Vectors 101: Pseudotyping Viral Vectors ... -
R Bodies: Membrane-Rupturing Microscopic Tools
TypeBlog PostPublishedApril 14, 2016, 2:30 p.m....muscle contractions) are produced by orchestrated movements of networks of proteins consuming molecular fuel...undergoing local conformational changes. These tiny movements are then amplified tens or hundreds of thousands...decades, and they were known to be resistant to detergents and degradation under a variety of conditions...see figure to the right), we identified R-body mutants that extend at both lower and higher pH. We initially...initially used this screen to find R body mutants that couldn’t extend at a normal pH and later realized...F. R. Organization and expression of genetic determinants for synthesis and assembly of type 51 R bodies... -
Hot Plasmids: Spring 2025
TypeBlog PostPublishedJune 3, 2025, 1:15 p.m....large library of iGluSnFR3 mutants, identifying the two best-performing variants as iGluSnFR4f (fast deactivation...iGluSnFR4f is well-suited for imaging rapid synaptic events such as in the somatosensory cortex, while iGluSnFR4s...display, they recommend the NGR vectors for most experiments. These new tools promise to be a popular choice...line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique...insertion through targeted CRISPR homing of retroelements." The Abudayyeh-Gootenberg Lab developed this...based on several new site-specific retrotransposon elements they identified and characterized (Fell et al....untranslated regions (UTRs) outside of the homology elements allowed for scarless insertion — no UTR sequence... -
Finding Your Perfect Job After University
TypeBlog PostPublishedJan. 12, 2016, 3:30 p.m....of the work, such as physically carrying out experiments like PCR and running gels, but in research you...in Ghana. My role involved teaching the nursing students and community health workers some basic biology...spreadsheets. It was great to be in the lab doing experiments without the constant fear of failure because...company carries out clinical trials of drugs and treatments for external companies. It was really exciting... particular situation, communication between departments was sometimes difficult and the solution attempted... companies which are reliant on contracts with clients is that, when those contracts fall through or are...pancreatic cancer where one drug does not work for all patients, leading to very high mortality. I really loved... -
Sweating the Small Stuff: Details in the Lab
TypeBlog PostPublishedFeb. 8, 2022, 2:15 p.m....small errors that significantly impacts your experiments. What do you do when you’re in the lab and you're... steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have a physical copy of...Have a physical copy on hand as you run your experiments. Try drawing out a protocol as you read it, or...an investment in accuracy. Bench setup Small adjustments in the physical organization of your workspace...you track samples as you add them. Moving your reagents from your left side to your right as you use them...are endless. Fig. 2: Blugene moves reagents from left to right as they use them. Prevent...focus, which was extremely effective. Others ran experiments at odd hours, used more secluded workspaces, ... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog PostPublishedFeb. 28, 2023, 2:15 p.m....consumer-grade components. In this field, often referred to as 'frugal science', existing components are adapted...opportunities) to modify the device for individual experiments. Manufacturers rarely provide the construction... by the manufacturers and enables customized experiments. Projects such as the OpenFlexure Microscope ... Taking this all together, the essential ingredients of open-source microscopy are as follows: Detailed...around the use of commercially available tools and components. Rapid prototyping techniques such as 3D printing...can literally put their work on the shoulders of giants. The still relatively young movement of sharing...similar goal. The sharing of modules, ideas, and blueprints has the potential not only to accelerate scientific... -
Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers
TypeBlog PostPublishedJuly 28, 2015, 2:30 p.m....of DNA Elements Project (ENCODE), an NIH-funded effort to define all of the functional elements in the...similar. There may be other variants of this that are better for some experiments than those we’ve put up....Eric Mendenhall of the University of Alabama in Huntsville explains it, a major goal in his laboratory is...groups got together to adapt CRISPR for tagging experiments and it’s already looking like it’s making a big... system is really quite simple but it offers improvements in efficiency and throughput. In adapting the...developed their own system or who use and adapt our reagents to think about adding them to Addgene’s special... -
Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
TypeBlog PostPublishedJune 2, 2015, 6:06 p.m.... just that as part of their CRISPR work in rice plants. Yang was intrigued by the tRNA processing machinery... in eukaryotes (E in bacteria), and only a few elements of tRNAs are necessary for recognition and cleavage...addition, tRNAs contain internal transcriptional elements that enhance pol III transcription, potentially...also examined editing efficiency in intact rice plants, comparing traditional gRNA constructs with PTGs... five gRNAs were tested, they found that 50% of plants had biallelic mutations at all five loci. This ...Although Xie et. al characterized the system in plants, PTGs will likely work well in many different biological... regions, such as noncoding RNAs or regulatory elements, to examine their function. Bacteria and viruses... -
Form Your Own Peer Mentoring Group: A How-To Guide for Scientists
TypeBlog PostPublishedFeb. 18, 2014, 4:22 p.m....Mentoring Group all participants act as both mentors and mentees. Because the participants are at a similar...make mentoring less mysterious and train the participants in skills useful for future mentoring relationships...at the same time Broadens the network of the participants Can be a lot of fun! Form your own peer mentoring...mentoring group In my travels speaking to grad students and postdocs I have started to talk to trainees....g. all women, all men, all postdocs, all grad students) or mixed for one or both of these criteria. If...you can, include people from different labs, departments or organizations as it brings a diversity of ...perhaps a 3 hour breakfast once a month. All participants must commit to making the meetings a scheduling... -
Giving gRNAs a Facelift - Synthetic and Beyond
TypeBlog PostPublishedOct. 20, 2022, 1:15 p.m....that stability of the RNA used in CRISPR-Cas9 experiments – the “gRNA” – suffers from the same issues as...nuclease-resistant, resulting in less off-target events. While both have similar advantages, bridged nucleic... Editing). This is particularly important in experiments in which viability is being compromised by introduction...CRISPR-Cas9 system can initiate targeted editing events. Both technologies utilize chemical modifications...gRNAs? Dye labeling the RNA in your targeting experiments allows you to visualize transfection efficiency...efficiency and even track localization in cellular experiments. There are many suitable dyes to choose from ...-703 (2020). https://pubs.acs.org/doi/10.1021/acscentsci.9b01093 More resources on addgene.org View ... -
Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards
TypeBlog PostPublishedAug. 5, 2021, 12:15 p.m....We are excited to announce the recipients of this year’s Michael Davidson and Roger Tsien Commemorative...researchers, in addition to postdocs and graduate students as we have done in the past. Below, read about...Filip Tomaska and Tim Hanson, are characterizing variants of iGluSnFR3 in vivo and have used it to detect...post-transfection, excited at 505 nm (right). Scalebar represents 100 µm. Image from Abhi Aggarwal. Brandán...providing insight into new methods for treating patients in recovery from opioid use disorder,” says Hubbard...line engineered to be electrically excitable. The mutants are co-expressed with a photoconvertible fluorescent...photoconverted, sorted, and sequenced to identify the mutants for further enrichment. Tian notes that this platform... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m.... from their native regulatory elements. Controls for RaPID experiments RaPID negative controls include...interest. See figure 1 below for details on the key components of RaPID. Ramanathan et al. also tweaked BirA... the role played by RNA-protein interactions in events that occur on a short timescale, such as during... BirA*-based affinity purification mass spec experiments. These background proteins were then subtracted...correlated with poor survival in breast cancer patients. By digging into the literature and analyzing ...disease. RaPID can also be used to test how RNA variants associated with human disease affect protein binding... with increased serum iron levels seen in HHCS patients. These are just three examples of how RaPID can... -
Human Germline Editing Using CRISPR
TypeBlog PostPublishedAug. 10, 2017, 2:19 p.m....Nature gives us even more to talk about, as it represents another step towards CRISPR germline editing ... the field will move forward from this work. Experiments & major findings Let’s start with the paper itself...forward with gRNA 1 to conduct microinjection experiments in S phase zygotes at 18 hr post fertilization...are needed for proper development. Ma et al.’s experiments showed a 3% mosaic rate, and this rate would ...to correct the hype when we see it. This paper represents a huge advancement in our understanding of how...perform in embryos but also shows that many more experiments are needed before we can consider clinical applications...applications for germline editing. Although this paper represents a large step forward in the CRISPR field, it’... -
The Future of Research Symposium Boston 2015
TypeBlog PostPublishedOct. 20, 2015, 2:30 p.m.... about postdocs. Among other involvements, he helped to establish grants through the Sloan foundation ...biomedical workforce. Quotes taken directly from participants (summarized in 2) touched upon many problems...hired to produce results, not scientists” These comments are well correlated with published data. For example...better solution would be to reduce the number of entrants into PhD programs to more closely reflect the ... market for PhDs Later this week, postdocs and students will again gather in Boston - this time with the...arguing against reducing the number of graduate students accepted in STEM fields (6). Melanie Sinche ...PMC4484856. 5. Bourne, Henry R. "A fair deal for PhD students and postdocs." Elife 2 (2013): e01139. Pubmed ... -
Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing
TypeBlog PostPublishedMay 5, 2015, 3:54 p.m....(electronic table of contents) alerts from journals you read often. The contents of each issue will be...explain some of the rationale behind the line of experiments, what data they collected and what it means. ... model or overall conclusion. In others, the experiments are geared towards eliminating possibilities ...deeper about the data they are showing. Are those experiments the best way of figuring out what’s going on?...controls? Refer back to the methods section for finer points of the protocols. Are there any red flags about... days of file cabinets filled to the brim with reprints and papers with scribbled notations in the margins...searching, reading and organization tips in the comments below. Additionally, read more of the Beginner... -
A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression
TypeBlog PostPublishedNov. 2, 2021, 1:15 p.m....Many neuroscience experiments that require gene expression in a specific cell type rely on transgenic...of off-target expression and ways to optimize experiments to minimize it. Overview of DIO/FLEx vector design...problem of DIO/FLEx vectors, the cause of these events, until recently, was not known. A recent paper ... both the serotype and promoter with in vivo experiments to determine the optimal/minimal injection titer...interest. You should now be able to make smaller adjustments to a set of injections into your animal model...may be tempting to skip some steps and get to experiments. However, by optimizing all conditions first,...results. Conclusion Using AAV vectors in your experiments requires a full understanding of vector performance... -
Popular Retroviral Vectors and Their Uses in Scientific Research
TypeBlog PostPublishedJuly 24, 2018, 12:51 p.m....). If you are wondering which virus fits you experiments best have a look at this viral vector overview...and expression of these vectors when used in experiments. Retroviral vectors make up a large toolbox ...screening of thousands of genes in genome-wide experiments. Gamma-retroviral vs. lentiviral vector systems.... More detailed descriptions of the lentiviral elements and the 2nd and 3rd Generation Lentiviral Systems...and tips for using retroviral vectors in your experiments. Gamma-retroviral vectors Popular gamma-Retroviral...can be used to induce site-specific recombination events and is widely used in the field of mouse transgenics...Zhang lab: We are constantly improving our CRISPR reagents. Please check www.genome-engineering.org for the... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m.... viral components or the transgene itself are used to quantify the number of integration events. Another...common assays for physical titer are direct p24 measurements via ELISA and qPCR for viral RNA. Measuring ...quantified using qPCR primers targeting specific viral components such as LTRs, gag, WPRE, antibiotic resistance-genes...genome and a cell that has had multiple integration events. In addition, promoter choice can also impact transgene... approach can discriminate multiple integration events, reducing the possibility of underestimating the...titering as this will better represent future experiments. The aforementioned titering options all have...of infection (MOI) for predicting gene transfer events." Genetic vaccines and therapy 2.1 (2004): 1. PubMed... -
The AAVantages of AAV in CRISPR Screens
TypeBlog PostPublishedFeb. 4, 2025, 2:15 p.m...., safety requirements and expression limitations may be too constraining for some experiments, particularly...that often affect which is chosen for certain experiments. AAV can be safely handled at BSL-1, while lentivirus...vectors is how the cargo — in this case, the CRISPR components — are expressed. Unlike lentivirus, cargo delivered...AAV genome can integrate at double-stranded break points caused by CRISPR-Cas9 editing. The good news is...the amount of cloning, another option is to use variants of Cas9, like split Cas9, or a smaller Cas9 ortholog...tissue-specific diseases has grown exponentially, aided by advancements in in vivo CRISPR screens. AAV vectors have ...have played a large role in these advancements, due to their many benefits for in vivo models. Over the past...