We narrowed to 570 results for: RON
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TypeBlog PostPublishedOct. 8, 2020, 1:15 p.m....sterilization as mycoplasma can even escape 0.2-micron filters. Figure 1: Contamination! Tissue culture...shortly. Once they are present in the laboratory environment, they are difficult to eradicate. You can suppress...maintenance of the cell lines. Providing such an environment for the lab facility will not only reduce mycoplasmas... you are careful enough to provide a sterile environment for work, it will soon become a habit before ...separate footwear for the lab to avoid bringing in environmental contaminants. Figure 2: An Addgenie...lines for assay or analytical purposes. Use 0.1-micron filters as are they are a better way of avoiding...avoiding mycoplasma, rather than using 0.2-micron filters, for media filtration. There is no 100% assurance...
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Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog PostPublishedSept. 3, 2019, 1:15 p.m....reversibly degrade proteins of interest fused to a degron, a domain that induces degradation. In plants, ...ligase complex, promoting its interaction with the degron sequence and ultimately its degradation. While ... needed for AID. By tagging proteins with a 7kD degron called mAID and expressing the essential plant ...channels that can be used to silence the activity of neurons. Earlier this year, the Hegemann lab and collaborators... cysteine. With the introduction of MerMAIDs, neurons can be turned off quickly and with high precision... a valuable optogenetics feature when studying neuronal activity. The researchers identified MerMAIDS ... -
Multiplex Genome Editing with CRISPR-Cpf1
TypeBlog PostPublishedMay 9, 2017, 2:12 p.m....there was strong expression of Cpf1 and GFP in the targeted brain region and ~75% of neurons were co-transduced...Cassettes transfection gRNAs expressed from a polycistronic transcript Requires co-transfection of Cas9...Cassettes transfection gRNAs expressed from a polycistronic transcript Requires co-transfection of Cas9 ...multiplex edit 3-4 genes in a primary culture of mouse neurons and in vivo. For these experiments, cells were ...-transduced with Cpf1 and GFP. Of these neurons, ~15% had indels at all 3 targeted loci. See the chart... -
Transferable Skills Guide: Public Speaking
TypeBlog PostPublishedApril 20, 2023, 4 p.m....Murphy’s law definitely applies: what can go wrong, will go wrong. The trash removal service slams dumpsters...useful transferable skill. Being able to stand up in front of an audience and convey your ideas clearly and...project. So whenever you freeze, or when things go wrong, ask yourself, “What next step will help me achieve...term “diving board” so you end up acting it out in front of a very confused group of visiting Chinese undergrads...incorrectly on slides 8-16. Your notes disappear. The wrong version of your slide deck gets uploaded. Your mic... -
Plasmids 101: The protein expression toolbox
TypeBlog PostPublishedFeb. 16, 2023, 7:05 p.m....Tet-on transgene vector. Degron tags Tagging relevant proteins with a degron tag can provide inducible...removed. Degron tag systems include small molecule assisted shut off (SMASh), auxin-inducible degron (AID)...SMASh degron tagging vector! Destabilizing domains Destabilizing domains (DD) are akin to degron tags,... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog PostPublishedJuly 18, 2019, 12:55 p.m.... RNA-sequencing to identify neuronal class and deep sequencing of neurons from each class to fully characterize...Addgene. C. elegans open resources The C. elegans Neuronal Gene Expression Map & Network (CeNGEN) The CeNGEN...comprehensive gene expression atlas of every C.elegans neuron. Scientists are currently doing this using two ...this system a modified TIR-1 protein can degrade degron-tagged proteins in an auxin dependent manner. This...Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed PMID: 30092212. PubMed... -
Viral Vectors 101: Voltage Indicators
TypeBlog PostPublishedJan. 30, 2024, 2:15 p.m....across the membrane of cardiac cells. Similarly, neuronal signaling is carried out by action potentials ...perceived signals to physical actions. Understanding neuronal pathways and the function of certain cells depends...be directed to specific types of cells, such as neuron subtypes, thanks to viral serotype specificity ...available VSDs has been successful even at the single neuron level in vitro. In vivo monitoring has also been...membrane as well. Another flavor of these tools is Voltron, which uses rhodopsin as the voltage sensing component...voltage is particularly physiologically relevant in neurons, myocytes, and endocrine cells. It follows that... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....norepinephrine (NE) expressing neurons and GFP (green) to label neurons transduced by each serotype. ...challenge to scientists. What works well in one cell or neuron type may not work well for a neighboring cell type...express ChR2(H134R) in A2 norepinephrine expressing neurons. We tested four serotypes containing an expression...84%) were both excellent at infecting our target neurons (Figure 1). However, this one set of experiments...Perspectives on Data Sharing in Deep Brain Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https... -
Getting to Know Expansion Microscopy
TypeBlog PostPublishedJune 20, 2023, 1:15 p.m....Chemical Synapses in the Brain. Neuron 68:843–856. https://doi.org/10.1016/j.neuron.2010.11.021 Gambarotto D...techniques like super-resolution microscopy or electron microscopy. Since its introduction in 2015 by ...2010). How do you overcome this resolution limit? Electron Microscopy (EM) is one option, but the equipment...helped researchers gain a deeper understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux... Ashley Waldron... -
Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More
TypeBlog PostPublishedAug. 13, 2019, 12:58 p.m....biological studies, including investigations into environmental pollutants and health, embryo growth, brain ...example, specific tissues such as blood cells or neurons can be fluorescently labeled, allowing researchers... for neuroscience investigations. For example, neuronal activity can be tracked in whole brains of swimming...high-resolution mapping of the brain by uniquely labeling neurons. Zebrafish are also amenable to manipulation of...consequences of activating or silencing specific neurons in living larvae. In addition, zebrafish have been...PubMed PMID: 18845710. Kim, Dal Hyung, et al. "Pan-neuronal calcium imaging with cellular resolution in freely... -
Five Popular Model Organisms, Part 2
TypeBlog PostPublishedSept. 29, 2020, 1:15 p.m....make them ideal for studying in a laboratory environment. We’ve covered five model organisms in a previous...candidate for introductory teaching and research environments (Idalia & Bernardo, 2017). Escherichia... disease-relevant genes in such a controlled environment displays how essential the frog is as a biomedical...genome, and is easily maintained in a laboratory environment. What sets Arabidopsis apart from other plant...Regeneration and Aging Research: A Mini-Review. Gerontology, 57(6), 565–571. https://doi.org/10.1159/000323761...the Axolotl Model for Regeneration and Aging. Gerontology, 66(3), 212–222. https://doi.org/10.1159/000504294... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog PostPublishedJan. 16, 2025, 3:42 p.m....Access to Projection Neurons. Neuron, 92(2), 372–382. https://doi.org/10.1016/j.neuron.2016.09.021 Walters..., 2023). It may be a good choice for targeting neurons, with high transduction frequencies in the central...to target the central nervous system, including neurons, AAV5 is generally the best choice for targeting... is derived from AAV2 and engineered to infect neurons and undergo retrograde transport, traveling up ... -
Antibodies 101: Validation
TypeBlog PostPublishedMarch 24, 2022, 1:15 p.m....molecule at all, leading to inconclusive results, erroneous conclusions, and wasted time. In fact, many have...recommendations for improving antibody use and reliability (Roncador, et al., 2015; Taussig, et al., 2018; Uhlen, et...that antibody works as expected, you will see a strong antibody signal in the retinal sample, but little... your antibody (B). Your results show a lovely, strong band from the cells transfected with your protein...Nature 518:27–9. https://doi.org/10.1038/518027a Roncador G, Engel P, Maestre L, Anderson AP, Cordell JL... Ashley Waldron... -
Hot Plasmids - October 2022
TypeBlog PostPublishedOct. 6, 2022, 1:15 p.m....mouse neurons. D) Membrane voltage traces of action potentials in a HcKCR1-expressing mouse neuron without...optogenetic silencer in experiments with mouse cortical neurons, far out-performing previous attempts to engineer...of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential frequencies with ...antibody anti-c-Myc [9E10] now available! by: Ashley Waldron The majority of Addgene's more than 50 antibodies... -
Three Tips for Preventing Viral Plasmid Recombination in Your Samples
TypeBlog PostPublishedSept. 14, 2021, 1:15 p.m....at Addgene, I get to see everything that can go wrong with a plasmid. One of the most frequent problems...smaller plasmids are formed. Viral vectors are prone to recombination In lentiviral vectors, recombination... elements. Knowing how to amplify recombination-prone plasmids and testing your prep can save you the ...H (2012) Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3...Genetic recombination of bacterial plasmid DNA: electron microscopic analysis of in vitro intramolecular... -
What the HEK?
TypeBlog PostPublishedApril 18, 2023, 1:15 p.m.... Frank Graham showed HEKs display many immature neuron-like qualities, which is inconsistent with kidney...is next to the kidney and is known to have some neuronal qualities.) While HEKs probably aren’t a great...researchers using the cell line. What the HEK is wrong with it? All cultured human cell lines are susceptible..., et al., Preferential transformation of human neuronal cells by human adenoviruses and the origin of ...Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021) (9):796991. DOI: 10.3389... -
Advice for Scientists Starting a Lab
TypeBlog PostPublishedApril 29, 2014, 1:43 p.m....equipment always helps. But how does one build a strong collaborative research team motivated by similar...your lab team and foster an effective working environment? Tom Ellis was offered a start-up package that...ways to improve the lab. “An effective working environment for research is totally dependent on communication...while others are more productive in a handsoff environment. “Treating people fairly does not mean equally...selectively picking the first couple of hires to build a strong, collaborative team. And take advantage of the ... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog PostPublishedNov. 6, 2018, 1:12 p.m....29] Photoreceptors [30] Otic hair cells [31] Neurons [32, 33, 34] You can find a variety of plasmids...have been successfully transdifferentiated into neurons and β-cells [40-42]. Reprogramming and transdifferentiation...S.C. Zhang, Directed differentiation of dopamine neurons from human pluripotent stem cells. Methods Mol ...human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat Protoc, 2012. 7(10): p...pluripotent stem cells to mature functional Purkinje neurons. Sci Rep, 2015. 5: p. 9232. PubMed PMID: 25782665...Direct conversion of fibroblasts to functional neurons by defined factors. Nature, 2010. 463(7284): p.... -
Targeted Mutagenesis with EvolvR
TypeBlog PostPublishedFeb. 21, 2019, 1:15 p.m....error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis with error prone PCR,...bacterial growth conditions. Expression of an error prone polymerase I (PolI3M) is capable of mutagenesis ...PCR, EvolvR uses an error prone polymerase to introduce mutations. However, by using nCas9 to direct PolI3m... Figure 1: EvolvR consists of an error-prone polymerase (PolI) that is fused to a nicking Cas9... -
A Quick Guide to a Career in Software Product Management
TypeBlog PostPublishedNov. 24, 2020, 2:15 p.m....skills in strong relationship-building and influence. You’ll need a large deal of influence and strong relationship-building...software product management was either. Coming from a strong background of undergraduate research straight to...energized from interacting with others, and have strong communication skills to tailor your message to ...implementation review. The developers discuss which front-end and back-end coding frameworks to use. I mostly... -
New and Upcoming Viral Vectors - May 2020
TypeBlog PostPublishedMay 28, 2020, 1:15 p.m....plasmid for expression in forebrain GABA-ergic interneurons (83895). See our Serotype Testing AAV inventory...with ever more efficient tools for light mediated neuronal control. The advantages of optogenetics lie in...system that allows for specific manipulation of neurons. We have expanded our AAV collection to include...targets expression of ChR2 to forebrain GAB-ergic interneurons through the mDlx enhancer element. Plasmid...DREADD acts through the Gq pathway and promotes neuronal firing. Plasmid Serotype Name Depositor 44361... -
Top Requested Lentivirus and AAV of 2016
TypeBlog PostPublishedJan. 6, 2017, 3:56 p.m....signaling pathways in specific cell types (typically neurons). While all of these viruses are popular, pAAV-...pAAV-hSyn-DIO-hM4D(Gi)-mCherry can be used to induce neuronal silencing in vitro and in vivo (Krashes et al..... The human synapsin promoter further specifies neuron-specific expression of the DREADD (Kügler et al...Lowell BB. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice. J Clin Invest..... Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression from an... -
RaPID Detection of RNA-protein Interactions
TypeBlog PostPublishedAug. 16, 2018, 12:40 p.m....increased iron serum levels and early-onset cataracts, is characterized by point mutations in the Iron Response.... In as little was one minute BASU generated a strong streptavidin signal (~250-fold enrichment) when...and IREB2, which post-transcriptionally regulate iron metabolism pathway transcripts. By using RaPID, ... loss of binding correlated with increased serum iron levels seen in HHCS patients. These are just three... -
New Tool for Lineage Tracing: The ClonTracer Library
TypeBlog PostPublishedSept. 22, 2015, 2:30 p.m....somewhat similar to an academic research environment” with a strong focus on publishing, but still made it...initial resistant populations can be eradicated up-front quickly. This approach is in contrast to the current...at cancer cell resistance to ABL1 inhibitors in chronic myeloid leukemia. Bhang says the work highlighted...2015): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct evolutionary... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog PostPublishedJune 26, 2018, 1:33 p.m....9514755. Richard Gronostajski: NFI-related diseases of neural development Richard Gronostajski is a professor...Paula Videira, Michael McMurray, and Richard Gronostajski who each are being awarded free plasmids form...they affect brain and muscle development. The Gronostajski lab has generated mice wherein it's possible...Campus in Aurora, Colorado, USA. Richard Gronostajski is a professor at the University of Buffalo.... -
Lab Automation at Addgene
TypeBlog PostPublishedMarch 22, 2018, 1:28 p.m....could ever do! However, there is quite a bit of upfront research you have to do to find said robot and ...into the robotics spirit include: Start using electronic multichannel and single channel pipettes. These...help keep things more organized. Cap tubes with a micronic cap sealer. This is a simple table top device ...has a roller to go over caps and push them into micronic tubes instead of pushing them in one by one. It... It seems very simplistic and it isn’t electronic but it is one of the most helpful machines I have used... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....antibody is produced from a new set of animals), and wrong application (different experiments/conditions can...that this antibody has specific binding and has a strong signal. We call the unique pattern of the antibody...7s) then the signal is weak and vice versa for a strong signal. However, exposure time and its relationship...compare exposure times of our samples. A prominent, strong signal has a shorter exposure time than the negative... neuroanatomy without the fear of referencing a wrong section. The antibody screening and development ... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog PostPublishedAug. 30, 2016, 2:30 p.m.... an extensive infrastructure of molecular and electronic companion technologies that has transformed the...CRISPR/Cas9-mediated genotyping, as well as our electronic reader that provides portable quantification ...Leslie Dan Faculty of Pharmacy at the University of Toronto. His research is focused on developing cell-free...biology applications for human health and the environment. Alexander A. Green is an Assistant professor...4. Messina, Jane P., et al. "Mapping global environmental suitability for Zika virus." Elife 5 (2016):... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog PostPublishedDec. 1, 2015, 3:30 p.m....conserved in mammals, expressed in neurons, and are especially abundant in neuronal synapses. shRNA silencing ...way to guide inheritance that might be less error prone than DNA replication. He too mentioned how “surprising...regulate at the DNA level and 1 at the RNA level. Udi Qimron from Tel Aviv University then spoke about using...treatment resulting in major problems in hospitals. Dr. Qimron proposes using bacterial phage vectors with CRISPR... -
PhD Applications After COVID
TypeBlog PostPublishedJan. 5, 2023, 2:15 p.m....interview is another way to make the virtual environment more comfortable and help you to keep yourself...institution, you can get a better sense of the environment of the department, the lab setting, and how students... can also be nerve wracking to ask questions in front of other prospective students; an online meeting...by talking to current students about the work environment and social aspects of the department, you may...Options Thanks to the much more developed virtual environment resulting from the pandemic, students who are... -
3D Printing Meets CRISPR Cas9
TypeBlog PostPublishedOct. 17, 2017, 12:55 p.m....images. In the early days of structural biology, Byron Rubin, a crystallographer working with Jane Richardson...solution to this problem with the development of Byron’s Bender – a mechanical device that could be used...protein’s structure into experimentally determined electron density, have been known to say something silly... in the education community that All models are wrong….Some models are useful. While a physical model ...first Cas9 model is certainly useful, it is also wrong in at least one serious way. We represented the ... -
Academic vs. Industry Postdocs
TypeBlog PostPublishedNov. 13, 2018, 1:22 p.m.... having already been immersed in an industry environment, have a better understanding of the fast, deadline-driven...up in an industry job, being immersed in that environment sooner rather than later can be helpful. Publishing...project might be challenging. In an academic environment, surrounded by lab members who are all working...Novartis, Biogen, Amgen, New England Biolabs, Regeneron, and Genentech, among others, and the list is ...opportunity to do some great science in a supportive environment. Both paths will give you myriad opportunities... -
Light Sheet Fluorescence Microscopy
TypeBlog PostPublishedJuly 16, 2015, 2:30 p.m.... In LSFM the laser light sheet, typically 2-6 microns, illuminates only one thin plane of the sample ...image acquisition is performed using cameras with electron multiplying couple-charged device (EMCCD) or complementary...of transgenic mice in which a subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H ...LSFM with multiple tissue clearing strategies and neuronal labeling methods will greatly aid our understanding...331–336. Pubmed. P. Keller, M.B. Ahrens. (2015) Neuron, 85, 462-483. Pubmed. E.H.K. Stelzer. (2015). Nature... -
Delivery Methods for Generating iPSCs
TypeBlog PostPublishedApril 17, 2018, 1:37 p.m....cloning capacity of ~8 kb, so either a single polycistronic vector or multiple single-gene vectors can be...endogenous pluripotency genes. Retroviral vectors are prone to incomplete silencing of reprogramming transgenes...differentiation potential. Finally, retroviruses are prone to insertional mutagenesis and random integration...retroviral vectors. This makes it possible to use a polycistronic cassette that expresses multiple reprogramming...factors can be delivered either as a single polycistronic transgene or with four different adenoviruses... ~14.5kb minicircle expressing a 2A peptide polycistronic cassette comprised of OCT4, SOX2, LIN28, NANOG... -
Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog PostPublishedSept. 3, 2020, 12:15 p.m....dimeric green fluorescent protein (pdDronpa) (Zhou et al., 2018). pdDronpa dimers dissociate in response to... to cyan light (500nm). By fusing the two pdDronpa domains to the sites flanking the DNA-binding cleft...binding in the dark. Upon light stimulation, the pdDronpa moieties move apart, allowing DNA binding and ...Cas9, dCas9 Gene editing Zhou et al., 2018 pdDronpa Cas9, dCas9 Gene editing, transcription Light-Dependent...photoactivatable transcription systems to induce neuronal differentiation. Nat Methods 14:963–966 . https... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m....dependent on the photon budget and pushing the frontiers of fluorescence microscopy often requires more...easier and more functional in complex biological environments such as live cells and tissues (Figure 1). These...several others can be made fluorogenic using environmentally sensitive fluorophores. Overall, self-labeling...photon budget and allow us to push further the frontiers of biological imaging. *Worried about your budget...Central PMCID: PMC4344395. 3. Lavis, Luke D., and Ronald T. Raines. "Bright ideas for chemical biology."... -
Lentiviral Vector Uses and Overview
TypeBlog PostPublishedMay 19, 2016, 2:30 p.m....system that could do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1...experimental use. In 1996, Naldini et al. transduced rat neurons in vivo with beta-gal and other reporters, and ...vectors are incredibly popular in the lab - the Trono lab’s lentiviral packaging and envelope plasmids...Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D. Science. 1996 Apr 12;272(5259):263-7. PubMed ...Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. J Virol. 1998 Nov;72(11):8463-71. ... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson & Bird, 2019) and is a global...of symptoms in animal models, Rett syndrome is a strong translational candidate for gene replacement therapy...therapy. In 2014, RSRT brought together a strong collaboration between two gene therapy labs (Brian Kaspar... half of the mutant MECP2 RNA in each of three neuronal subtypes of the hippocampus. This technology has...Modifications in Human-Induced Pluripotent Stem Cells. Front Genet 10: . https://doi.org/10.3389/fgene.2019.00625... -
Career Insights: Technical Support Specialist
TypeBlog PostPublishedNov. 9, 2017, 2 p.m....myself in a technical support role only when I was confronted one day with a product issue in the lab. The ...the world easier. You should also demonstrate a strong willingness to help others with their problems,...sometimes comprised of several layers of support: A front-line team accepts any incoming requests and answers...because there are just so many things which can go wrong in a lengthy protocol. And customers may tell all...careers are open to experienced professionals with strong interest in the technical aspects of science and... -
Supporting Reproducibility with a Connected ELN
TypeBlog PostPublishedJan. 17, 2019, 1:38 p.m....virtue of liberating data from paper into an electronic environment and hence making it searchable and shareable...founder of Research Space. There are many types of electronic lab notebooks (ELNs), each with its own pros ... of Research Space, which provides the RSpace electronic lab notebook. Rory is based in Cambridge, Massachusetts...on the Addgene blog Find tips on choosing an electronic lab notebook Watch a livestream of the Minisymposium...