..., an individual experiences progressive muscle wasting leading to death at around 30 years of age. Despite...the amount of research conducted on DMD, there is still no good treatment. The dystrophin gene is very large...approach. Since shorter forms of dystrophin can still be functional, exon skipping is a good option for...non-homologous end joining (NHEJ) pathway. It’s estimated that very little dystrophin correction (about ...could make a huge difference in DMD, and it’s estimated that exon skipping therapies would be applicable...experiment, they found the exon-skipped transcript constituted 59% of total dystrophin mRNA, similar to the ...CRISPR therapy closer to the clinic, much work is still needed. First, CRISPR delivery must be optimized...

... preparations undergo rigorous quality control testing at Addgene meaning they come ready made to accelerate...researchers lamenting the viral production process while still ordering viral vectors. Virus production can be ...interested in faster by producing, titering, and testing viral preps for them. At the same time, we’re developing...base for the processes of viral production and testing so that researchers can use our educational resources...researcher Connie Cepko has been helping us with AAV testing and recently had this to say about the service:...suggest additional plasmids with the Viral Service Suggestion Form. Lentivirus As some of the most popular ...well as other expression data. If you have any questions about the Viral Service or any of the materials...

...to non-jargon in the context of disease testing, it can still be considered jargon in most other uses....situation rather quickly!  The question   The first, and most important question, you should ask for any science...ll see throughout the posts and introduce *the* question underlying all communications efforts.  Scicomm...scientific or science communication. A Tweet is still considered scientific communication if the language...An audience can be defined by any number of characteristics. Common ones include age range, familiarity...patients or caregivers affected by the disease in question who are involved in online patient/caregiver communities...

...action and could potentially stimulate the development of useful diagnostic methods and improved therapies...for PZA-resistant Mycobacteria tuberculosis. Investigation of multispecies biofilm formed by Pseudomonas...compounds on these proteins will be tested. Investigating changes in neuronal activity after ischemic ...ischemic brain injury Dr. Gohar Tsakanova, Institute of Molecular Biology of the NationalAcademy of Sciences ...Armenia Gohar Tsakanova is a Senior Researcher who investigates the molecular and cellular etiopathomechanisms...non-linear microscopy methods will be used to stimulate and manipulate neural populations in order to ... Julio Ernesto Valdivia Silva is a master of interesting combinations: biomedicine and astrobiology. During...

...adjust our estimates based on the journal that published the articles.  That is, we re-estimate the citations...correlation. But even with that caveat, it is still suggestive that we see such a big difference in citations...than those without Addgene deposits? Even more interestingly, could we tell if depositing a plasmid with ... This is also true for other journals, so our estimates now take into account any differences in citations... That papers with plasmids deposited at Addgene still accrue on average 2 to 3 times as many citations...step. There are lots of other factors that could still be controlled for. As an example, plasmid-related...

...experiments.” - Alex Chavez, researcher at the Wyss Institute for Biologically Inspired Engineering and member...alleviate this frustration by producing, titering, testing, and providing lentiviral preps of select lentiviral.... For example, we’ve teamed up with the Broad Institute to provide lentiviruses for delivery of SpCas9...they require extensive background work from the requesting scientist for their proper use; after receiving...already expressing Cas9. As John Doench (Broad Institute and member of the Addgene Advisory Board) says...about using our viral services. Please send any questions or comments to [email protected]. Finally we’d ...and, in particular, the Church lab at the Wyss Institute for Biologically Inspired Engineering and the ...

...Hernández-Sánchez is a postdoctoral researcher at Instituto de Investigación Biomédica de Salamanca (IBSAL) and one...the competitiveness of cutting-edge research, is still one of the most valuable features of modern science...researchers, such as postdocs and principal investigators. They also come from many different scientific...-and-error. A recommendation for the correct statistical test can save a retraction. Today, collaborations... favors from becoming starting points for long-lasting partnerships Join us! We are a community of researchers...about us and what we do in our Frequently Asked Questions. We also invite you to participate in the 1st ...16:00 CEST. This symposium aims to answer all questions you may have about Crowdfight and we will have...

...majority of research institutes.  Pro tip! In most academic and nonprofit institutes, the Tech Transfer ...our process easier for others.  If you’ve still got questions about our MTA process, you can check out ...commonly used ones for transfers between academic institutions.  What is the UBMTA, you ask? Well, organizations...people think our MTAs are different than other institutes. But they’re not! The vast majority of our plasmids...’s important to note that it’s the depositing institute that decides the terms of distribution for their...of coordinating signatures from two different institutes. Our online portal automates a lot of the process...

...Addgene Advisory Board member, and Institute Scientist at the Broad Institute, John Doench. CRISPR technology...nucleotide transitions but still requires a nearby PAM, although these are still early days for this technology...gRNAs to 5 – 65% of the protein coding region will still result in many dozens of gRNAs to choose from. With... While SpCas9, with a PAM preference of NGG, is still the most widely-used Cas enzyme, the development...basic landscape of mismatches that can nevertheless still lead to activity has been established, and can be... different sequences targeting the same gene, suggesting that off-target effects did not overwhelm true...the Genetic Perturbation Platform at the Broad Institute and has worked with many Addgenies to help improve...

...maintain fidelity with the stimulus. For example, if you want to stimulate at a fast frequency without...hardware types, construction, and testing used for delivering your light stimulus. This can make or break your...that induce conformational changes with light stimulation to either activate or inhibit a specific receptor...cryptochrome 2 (CRY2) (Zhang & Cui, 2015). When stimulated with light, the endogenous proteins activate ...slower kinetics can be used for lower frequency stimulation. GPCR signaling GPCR signaling is a complicated...be very different responses to long vs. short stimulation times or frequencies (Zhang & Cui, 2015). You...Bendifallah, I., Zhou, F., Zeitzschel, N., Ahlbeck, J., Augustin, S., Sauter, K., Papagiakoumou, E., Gottschalk...

...someone asks a question about something they did not understand about your presentation, question whether other...beneficial to pose these why questions to yourself, but ultimately there is no substitute for feedback from others...research proposal is the perfect fit to their institutional focus and that you're better equipped than anyone...audience think or act.   To address that 'why' question effectively, you need to make your audience the...communication should prepare you to ask and answer these questions for a diverse set of audiences.  Every scientist...opportunities to practice communicating.  Some fantastic K12 education initiatives like BiteScis (https...

... social media conversations, conferences, and questions our scientific support staff receives – in fact...and share in-depth answers to frequently asked questions. We also want to tell you about things you didn...articles answering all of our frequently asked questions!    Identify needed content Let’s take a look...successfully start using them at the bench, along with questions we thought they might have as they go. The guide...things and another for asking specific science questions. It’s not uncommon to see requests for fact checks...of our content already comes from you and the questions you ask Addgene and your research community! We...engaging and informative resources that answer questions our depositors and customers are asking. So how...

...blot *indicates a diagnostic-only step Issue Potential Cause Troubleshooting Suggestion Proteins are...done the technical design of your blot, but are still finding yourself having issues, you’re in the right...type of gradient can help answer your particular question before running one.  Good on gradients? Excellent...the end of the post) has a number of helpful suggestions.  Gels Gel issues usually come down to voltage...want to try optimizing your transfer buffer.  Adjusting the buffer For wet and semi-dry transfers, you...membrane while increasing the ability of proteins to stick to the membrane by stripping away the SDS. To speed...blocking conditions on the spec sheet. If you’re still seeing high background after following manufacturer...

...contributed by guest blogger, Eugenia Rojas. A question worthy of a PhD: How do you visualize protein ...or in any other cell type. However, we cannot distinguish new from old proteins within that pool. Therefore...were quite tedious and, even here, there is the question of whether or not the tags influence behavior.... international PhD students at the Max Planck Institute for Biophysical Chemistry, Göttingen in Germany...in situ. I was in luck when I attended a very interesting talk by Prof. Erin M Schuman, a former Caltech...Caltech researcher, now Director at the Max Planck Institute for Brain Research in Frankfurt am Main, Germany...Germany. She presented a novel technology, back then still unpublished, that allowed the visualization of specific...

...The Rare Genomics Institute BeHEARD (Helping Empower and Accelerate Research Discoveries) Challenge provides...our fourth year working with the Rare Genomics Institute by providing plasmid-based tools for rare disease...developmental delays or autism spectrum disorder. It is estimated that CTD is the second cause of male intellectual...psoriasis is a chronic autoinflammatory skin disease estimated to affect 0.27 to 4.6 per 10,000 people globally...-36 receptor inhibitors in development, there’s still a lot of mysteries surrounding IL-36 biology. “Little... the signalosome of different receptors upon stimulation. “To study cellular transduction events, we will...would allow us to address many of our research questions,” Hébert says. In the spirit of plasmid sharing...

...DEPC.  Ensure that plasticware or glassware used is RNase free. RNase-free plasticware is readily available...doing something without a kit has advantages: less plastic waste, less expense, and less of being left with...product to get rid of RNases such as RNaseZAP. When harvesting tissues, cells, plants, fungi, or bacteria, keep...extraction methods evolved into a simple protocol still used today There are many alternative methods for...-phenol-chloroform. In this method, samples are still homogenized and lysed in a guanidinium thiocyanate...hours, and variations on this no-kit method are still widely used today (Chomcynski and Sacchi, 2006)....Making the simple protocol even more foolproof (still without a kit!) As mentioned above, working with...

... “designer” channelrhodopsins with desired characteristics. The model was trained to predict if a channelrhodopsin...stronger photocurrents at low light intensity than existing channelrhodopsins CoChR and/or ChR2(H134R). Designer...however, have a higher conductance, so they can still depolarize neurons at lower MOIs. Figure ...located in the midbrain, fiber-optic cannulas were still stereotaxically implanted into the mice’s brains...reach the VTA. Mice were then allowed to “self-stimulate” their VTA dopaminergic neurons by triggering ...H134R) expressing mice did not have this self-stimulation response. This result showed that ChRger2 stronger... systemic delivery with PHP.eB compared to the existing ChR2(H134R).  In the second experiment, ChRger2...

...Drugs, DREADDs) and optogenetics as tools to investigate the roles of certain cell types in locomotion...themselves are not pathogenic, precautions should still be taken when using them. There are suspected cases...cause the least amount of immune response. Any investigator who is using an in vivo animal model should ... useful for a wide variety of experiments, care still needs to be taken when selecting which particular...do you choose the right serotype? Keep asking questions!  Table 1: AAV Serotypes and Tropism Experiment...transport, but they also have similar transport characteristics (velocities of movement, distance traveled,...excitatory connections (3). These facts beg the question: is axonal transport desirable or not desirable...

...Biotechnology from Worcester Polytechnic Institute where he investigated how plants achieve polarized cell growth... and Biotechnology from Worcester Polytechnic Institute. What is RNAi? The loss-of-function (LOF) experiment...any technique in the context of the biological question. For example, while gene knockouts have long been..., gene knockout strategies cannot be used to investigate essential genes. Historically, gene silencing...work has called this prevalent assumption into question. Specifically, CRISPR-generated mutants believed...present at the first stage of an RNAi-based investigation. Creation of the RNAi trigger requires a few...This process, known as “transitivity,” calls into question any claims about specificity of a given RNAi technique...

...and optogenetics meet, allowing scientists to investigate the fundamental basis of both brain function ...combined optical and genetic methods to photo-stimulate mammalian neurons and alter synaptic events: a...rhodopsins and neurons have in common? Neuronal stimulations are triggered by the movement of ions across...viral-infected neurons are subsequently photo-stimulated through fiberoptic cannulas directly implanted.... In particular, they found that optogenetic stimulations of the DA neurons produce behavioral and electroencephalography...could activate appetitive rather than aversive stimuli. In a recent paper, de Jong and colleagues used...used rAAV FLEx-based vectors to answer this question, mapping and characterizing the activity of the DA ...