We narrowed to 807 results for: APP
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TypeBlog PostPublishedApril 24, 2015, 2:39 p.m.... Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing in other systems...Between Brainbow Systems Brainbow variants and applications beyond the mouse brain In addition to improving... Currently, Brainbow techniques are also being applied to study model organisms such as Drosophila and...zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as connections between...Brainbow’s colorful labeling has decreased the mapping time for a given section of brain by at least an...brain and other biological systems. Interested in applying Brainbow to your research? Check out Brainbow ... Nov 1;450(7166):56-62. PubMed. A technicolour approach to the connectome. Lichtman JW, Livet J, Sanes...
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Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog PostPublishedSept. 3, 2020, 12:15 p.m....improvements to CRISPR technology since its initial applications for genome engineering in 2012. Many of these...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed...allowing DNA binding and cleavage. This technique was applied to create photoswitchable spCas9, dCas9, and S....saCas9) proteins, showing the versatility of this approach. This study generated the first optically active..., allowing the release of Cas9. A noteworthy application of the CASANOVA system was its use in light-mediated...Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically encoded ...Photoactivatable moiety Cas9 variatn CRISPR protein application Split Systems / Light-induced Dimerization ... -
Deep Dive: qPCR
TypeBlog PostPublishedSept. 29, 2022, 1:15 p.m....understanding and technical proficiency with common applications - now let’s dive right in! You know qPCR is...as it solves, so understanding and using the application correctly is imperative. Picking your chemistry...space. For many cell lines and tissue types, appropriate reference genes (and supporting data) are often...2002), though there are cases in which it is an appropriate choice (Panina, 2018). Look for studies specifically...Vandesompele, 2002) will help you select the appropriate reference genes - and it is well worth your time... more, take a look at their 551 page e-book on applying MIQE guidelines! With these kinds of resources...melting profiles of PCR products in a rich web application, Bioinformatics, Volume 27, Issue 7, 1 April ... -
Bringing Sustainable Practices to the Lab: Innovations
TypeBlog PostPublishedMarch 7, 2019, 2:39 p.m....New ideas and approaches are encouraged in science. It isn’t surprising that new ideas are also popping...lab lives. Read on to learn about some creative approaches to lab sustainability, and find ways to further...but what about all the manufacturing steps that happen before you even receive the product? How can we...tips. This is part of a cultural change in how we approach waste that needs to be supported at all levels... facility, where it’s sanitized through an EPA-approved process and no longer considered hazardous. Then...industrial and residential markets. Rethinking our approach to lab waste is something we all need to do. Remember... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog PostPublishedFeb. 13, 2025, 2:15 p.m....Early CRISPR applications were often limited by the low editing efficiency of homology-directed repair...BE3, nicks the unmodified DNA strand so that it appears “newly synthesized” to the cell. Thus, the cell...reduce the undesired C→G or C→A conversions that can happen with BE3s. These byproducts likely resulted from...making them a promising tool for cell therapy applications. Dual base editors What about combining the ...Grünewald et al., 2020). Another lab took a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion...activity compared to ABEmax. In an alternative approach, engineering and directed evolution of the TadA...editors with increased activity and therapeutic application. Nature Biotechnology, 38(7), 892–900. https:... -
Antibodies 101: The Basics of Western Blotting
TypeBlog PostPublishedMarch 8, 2021, 2:41 p.m....of the protein of interest. An electric current applied to the gel causes the negatively charged proteins...with blotting paper and an electric current is applied causing proteins to migrate out of the gel and ...sandwiches. Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa. In addition to validated applications, pay close attention to the controls listed ...choose antibodies that are validated for your application and test the antibody in your specific system...to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences... -
Recombinase-based State Machines Enable Order-dependent Logic in vivo
TypeBlog PostPublishedJuly 28, 2016, 2:30 p.m....attB-attP pairs from different recombinases are overlapping or nested on a register, then the operation of...more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The re-arrangement...performed on one register at a time. If a particular application makes it difficult to isolate single registers...state read-out method may not be suitable for applications that require minimal cellular perturbation. ...contains the register. Depending on the desired application, the inducible promoters we used to drive recombinase...program to find the one that best suits your application. How efficiently do RSMs work? We tested a two-input...anticipate that, depending on the context and application, some inducible recombinase systems may work ... -
We're Updating Our Hold for Publication Policy
TypeBlog PostPublishedJune 24, 2025, 1:15 p.m.... complete necessary quality control and legal approvals so the plasmids are ready for distribution as ...this Hold for Publication policy seriously and appreciate the trust you’ve put in us when you choose this...worry, we’ll let you know before these releases happen (and congratulate you on your paper!). And as always...your deposit or plasmid ID numbers and we’ll be happy to update the plasmids to add the citation and release...hope somebody will find the plasmids useful, we’re happy to distribute unpublished materials. What about ...needs, so please let us know what you think. We appreciate your feedback. If you have questions about depositing... -
Year of the Camelid: Antibody Style
TypeBlog PostPublishedMay 9, 2024, 1:15 p.m....diagnostic applications. For example, fluorescently and radioactively labeled sdAbs have become appealing candidates...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative... as well. The first sdAb based therapeutic was approved in 2018, with many others in clinical trials (...maintaining a llama facility is out of reach and unappealing to many labs), the field is moving towards using...inspired this versatile family of tools, the applications they have already enabled, and the discoveries... -
Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi
TypeBlog PostPublishedApril 25, 2019, 3:01 p.m.... be used as a fusion tag for a wide range of applications from traditional imaging to FRET and stochastic...transforming E. coli with a gene library of approximately 484 variants, including some potential tetramer-breaking...Florida State University, they used a mutagenesis approach and found that specific amino acid changes favored...violet range. For example, mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not...fluorescent protein: structural characterization and applications in fluorescence imaging." Biochemical Journal...identification of the molecular determinants of color and applications in fluorescence imaging." BMC biology 6.1 (2008... -
Important Considerations When Using AAVs
TypeBlog PostPublishedJune 13, 2017, 2:30 p.m....possible tools, lentiviruses for example, are not approved for use in vivo use at the University of Manitoba... be mostly on the brain, but many of these tips apply to all tissues. Subscribe to Viral Vector Blog Posts... infect) can influence an AAV’s value greatly. Applying this mantra to AAV selection, there are three ...of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD complexes...only can we compare these methods in specific applications, we may also be able to use these comparisons... on overall cell function. AAVs are the most appropriate vehicle for this type of research, as they are...efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities. Many... -
9 tips for a successful postdoctoral experience
TypeBlog PostPublishedOct. 9, 2018, 12:21 p.m.... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia... have been completed. Therefore, to be ready to apply for a Group Leader position, key first author manuscripts... project. Furthermore, Group Leader position applications require 3 letters of reference- the graduate...informal recommendations, more positive grant application and manuscript reviews, etc. You'll also meet... fluorescent proteins, and live cell imaging approaches. Erik is also an avid long distance runner, gardener... -
"Build Your Plasmid": The Game - Play to Teach
TypeBlog PostPublishedJuly 17, 2018, 12:23 p.m....information they’ve absorbed if they don’t get a chance apply it in an interactive setting. The problem is…how... as I try being compelling, interactive, and application-driven to boost curiosity, I have never experienced...they would use their logic and strategy skills to apply those concepts and understand how therapeutic strategies... BUILD YOUR PLASMID is a game where you need to apply basic molecular biology, cell biology, microbiology...best strategy and propose a final gene therapy approach for the disease to be openly discussed with the...opposing team. In the end, the students had fun while applying most of the concepts required by the course and... -
Science Career Options
TypeBlog PostPublishedFeb. 23, 2017, 1:30 p.m....critically and carry out in-depth research is applicable to a wide range of industries and is a highly...skills acquired from a science degree that can be applied to many roles. These types of skills show that ...sectors such as forensics currently have too many applicants compared to the number of available roles). Make... Make your resume stand out Before you begin to apply for a job, it is useful to search online job websites...your research, it’s time to start sending out applications. Check out our other career posts for more specifics...careers and to find advice on crafting the perfect application. Additional Resources on the Addgene Blog Learn... -
Predicting Adverse Reactions to Monoclonal Antibody Drugs
TypeBlog PostPublishedFeb. 13, 2024, 2:15 p.m.... Limitations of the in silico approach The in silico approach cannot replicate the complexity of ...Better together Combining in silico and in vitro approaches enables researchers to develop safer and more...unwanted immunogenicity. The 3-D models can be applied to corroborate the occurrence and extent of the... somewhat act like a 2-step verification. All approaches have their own limitations. Using methods together...together can inform decisions about in vivo application and clinical testing. This would pave the way for... -
Management for Scientists: Delegating is Key
TypeBlog PostPublishedFeb. 20, 2015, 8:30 p.m....dramatically improved when tasks are distributed appropriately. If team members have opportunities to develop...develop and stretch then they will be happier and more productive in the long run. If you are overwhelmed...the copy, design every icon, curate all of the appropriate plasmids, and code the entire page. That would... page. The team manager must then facilitate appropriate communication between members of the team to ...some important criteria for delegating tasks appropriately: Are the desired goals and outcomes clear? ...questions. If you are perceived as too busy to be approachable mistakes will be made that could have been prevented... -
AAV Q&A with Tim Miles
TypeBlog PostPublishedMay 17, 2022, 1:15 p.m.... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be... be best served by a dual vector, split intein approach. Q3: What’s the best method for titering AAV...1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011 - 1x1012 vg/...systemic injections? A: We have not observed any appreciable difference in AAV transduction efficiency between...CRISPR tools is an exciting area for future AAV application to epigenetic editing. This area is discussed... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....editing applications. Figure 4: Overview of CasMINI for genome engineering applications. Image ...stability in cells, and use it in a variety of applications. This set of new anti-FLAG frankebodies includes...to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling various targets... and anti-HA frankenbodies have a variety of applications, including live-cell imaging, single-molecule...challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature CRISPR-Cas system ... -
Viral Vectors 101: Types of viruses
TypeBlog PostPublishedJune 6, 2023, 1:15 p.m....cell type doesn’t always lend itself to direct application of virus. However, the cargo capacity of AAV ...gamma-retro viruses. This is large enough for many applications, but in some cases multiple AAVs (co-infection...and AAV. If you’re wondering more about what applications these viruses are good for, we have plenty of...Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New...Gammaretroviral vectors: biology, technology and application. Viruses. 2011 Jun;3(6):677-713. DOI: 10.3390... Campos MD, Materatski P. An Overview of the Application of Viruses to Biotechnology. Viruses. 2021 Oct... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog PostPublishedJune 2, 2016, 2:30 p.m....information in the future. Approach: Outline a brief overview of the approach that you will take to help...biologists are finding themselves using computational approaches to analyze large data sets (myself included) ...your analysis, even if your conclusion is “this approach is not ideal because…” Including a conclusion ...or may not be where you also record your goal, approach, and conclusions (see above). Keep original ...in my own research. Again, some of this may not apply or may not work for you. The best way to find a ... -
Antibodies 101: Designing Your First Flow Panel
TypeBlog PostPublishedMay 28, 2024, 1:45 p.m....fluorophores, together, are able to give you the appropriate readout. A common method of labeling proteins... wavelength. Likewise, the emitted photons also appear within a range of wavelengths. For that reason,...excitation and emission are the most efficient (i.e., happen most of the time). Figure 1 shows a graph from ...higher-wavelength emission spectrum compared to APC. This happens due to a process called Fluorescence Resonance ... the fluorophore, and whether that fluorophore appears alone or in conjugation with another fluorophore...absorption at 650 nm, APC-Cy7's optimal excitation happens at higher wavelengths of about 750 nm. Source: ...Besides some background cell death constantly happening in cell culture, the transduction process itself... -
A Primer on Optogenetics: Introduction and Opsin Delivery
TypeBlog PostPublishedSept. 10, 2015, 2:30 p.m....upcoming posts in this optogenetics primer series, I’m happy to share some of the tips and tricks and I’ve learned...memory task, on the other hand, would not be an appropriate test. Of course, proper controls are required...using these lines is, if one is crossed with an appropriate mouse line expressing a particular cre, viral...22430017. 2. Fenno L, et al. The development and application of optogenetics. Annual review of neuroscience...PMC3225502. 4. Mattis J, et al. Principles for applying optogenetic tools derived from direct comparative...neuronal populations using combinatorial viral approaches. Current protocols in neuroscience / editorial...PMC3191888. 9. Zeng H, Madisen L. Mouse transgenic approaches in optogenetics. Progress in brain research. ... -
Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools
TypeBlog PostPublishedApril 23, 2018, 2 p.m....developed by the James and Schuh labs. Clift et al. applied TRIM-away to nine proteins in ten ddifferent cell... sometimes incorporating chemically inducible approaches to provide temporal control, this new system,...Epigenome editing by Cas9 (FIRE–Cas9), combines these approaches to enable rapid and reversible recruitment of...target multiple loci at once. In addition, the approach allows minute-by-minute analysis of the kinetics...domain. Transcription factor activation via the appropriate ligand - estradiol for hER or progesterone for...leads to nuclear localization, binding to the appropriate promoter sequence and activation of gene expression... -
Meet an Addgenie: Insights into Grad School, Career Paths, and More
TypeBlog PostPublishedJune 25, 2020, 1:15 p.m....the co-op application process, my advisor encouraged me to keep my values in mind as I applied. Thinking...My name is Lauren Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst...s and leave. I was really devastated when that happened. I didn't know if I should even stay in science...in your career path and still find comfort and happiness there. Read all of the Meet an Addgenies features...connections with my coworkers that may not have happened if we hadn’t had these conversations. Now, I am... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog PostPublishedMarch 12, 2020, 1:15 p.m.... Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells. To do...type specific expression in the retina. Four approaches for designing synthetic promoters To build a...reproducibly labeled retinal cells can be found here. Applications for synthetic promoter AAVs Besides restricting...synthetic promoters have a few other potential applications: Using promoter combinations for targeted expression... promoters expressed in. The lab tested this application using one promoter to drive expression of Cre-mCherry... rates of labeling are ideal of gene therapy applications, promoters that produce sparse labeling would... -
Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation
TypeBlog PostPublishedDec. 14, 2021, 2:15 p.m....simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ...and Szyf 2021). Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...resistance genes. It’s that simple! Good luck and happy editing! Thank you to our guest blogger!Daniel...Addgene blog CRISPR101: Editing the Epigenome Mapping the 4D Nucleome with CRISPR/Cas9 Resources at ...Xu X, Tao Y, Gao X et al. (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discov 2,... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m.... to increase AAV transduction efficiency by approximately 15-fold (Chen et al., 2006). 3. Transduction... at 4 °C). After ultracentrifugation, virus may appear as a white pellet. The pellet can then be rinsed... This antibiotic selection can be performed approximately 72 hours after transfection and should give ...give rise to a surviving population of approximately 20–50% of the packaging cells after 1–3 days of selection... could be examined with a microscope and should appear fluorescent if they were infected. While there ...transfection, higher-level expression after approximately 72 hours is likely the result of successful ... -
Tips for Titering Your Lentiviral Preps
TypeBlog PostPublishedMarch 15, 2016, 2:30 p.m....to be less expensive than an ELISA kit. In this approach, viral RNA is first converted to cDNA and then...several different lentiviruses in parallel, this approach should be considered. Similar to the p24 assay... Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried...have integrated into a target cell’s genome. This approach can discriminate multiple integration events, ...underestimating the viral titer. The main drawback of this approach is that it can be quite labor intensive. Target...virus may want to choose more straightforward approaches such as the FACS-based or colony-forming unit... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog PostPublishedDec. 10, 2020, 1 p.m....18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair...phenomenon to the presence of terminal proteins capping the ends of linear double-stranded AdV genomes....double-stranded chromosomal DNA breaks “that always happen naturally.” These adenoviral CRISPR/Cas9 genome...and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral vectors Since... Adv vector delivery is often used, the CRISPR approach based on HDR has the advantage of being a potentially...Strategies for Tissue-Specific Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp ... -
Of Myc and Men
TypeBlog PostPublishedJan. 19, 2023, 2:15 p.m....The strategy they landed on was to recombinantly append their proteins of interest with unrelated peptides...generate a reliable antibody for protein tagging approaches” was not part of the goal when Evan’s team generated...plays in human health and disease. But no science happens in a vacuum! Munro and Pelham learned of these... flexible - it could now be used in way more applications. Fig. 1 Making a Protein Tag (Abridged...their study, some of the newer Myc-tag antibodies appeared to be less sensitive to sequence context. So...proteins expressed from cloned genes: deletion mapping functional domains of Drosophila hsp 70. Embo J...