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...improved, by adding a nuclear localization sequence (SV40 Large T nuclear localization sequence) and expressing...asymmetric target site core but its 8 bp length that is critical for Cre and FLP function. Because recombination...Recombination between the two target sites with identical sequence leads to inversion of the flanked DNA...this plasmid mix results in sparse labeling of cortical neurons and their synapses.   How can you...control   Figure 2: Expression of a morphological marker (eYFP) and synaptic marker (PSD95-mcherry...

...DnaA binds multiple specific sites within the ori called DnaA boxes. These boxes come in both high- and ...Created with BioRender.com. Once DnaA breaks the local hydrogen bonds holding the DNA together, it recruits...itself. RepA binds to directly repeated sequences called iterons located near the ori and directly interacts... the protein to be translated. An antisense RNA called Inc, also encoded by the ColIb-P9 plasmid, is complementary...between iterons on different plasmids in a process called "handcuffing," which blocks replication. ColIb-...Replication Browse other Plasmids 101 Posts Get practical molecular biology help with our Molecular Biology...

...asbestiform minerals such as erionite, are the only scientifically proven causes of the disease. There is no question... question on this point within the medical and scientific communities, and the link between asbestos and.... New techniques are currently being tested in clinical trials to detect mesothelioma earlier than ever...the high mobility group box 1 protein. Normally localized to the nucleus, if the cell dies from injury, ...brought a lot of promise to a field that has historically been full of bad news and dreadful prognoses...Study Cancer Drug Resistance Plasmid Grant to Aid Cervical Cancer Screening in Ghana Additional Resources...

...defined chunks, called stories, and work on stories during short periods of time called sprints. The team...code, we might be calling something a sample and in other places, we might be calling it an aliquot. This...follow an agile methodology of software development called Scrum. The Scrum framework is based on an iterative...ourselves. We are also able to consider different technical approaches that may come up during discussion....

...cryogenic electron microscopy (cryo-EM) protocols typically require high concentration and purity of the target...limitations, the Funabiki Lab developed a technique called Magnetic Isolation and Concentration (MagIC)-cryo-EM...circles), magnetic bead (*), and spacers are labeled; scale bar: 100 nm. D) 3D model of the obtained structure...right). B) Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of ...orientations, this could be overcome with methods such as chemical crosslinking or tilted-stage imaging. A truncated...evolved recombinases and prime editing. Nature Biomedical Engineering, 10.1038/s41551-024-01227-1. doi:...

...combining these two strategies allows you to specifically incorporate homo- or heterozygous mutations....in the field at the time but had never been systematically analyzed: introducing silent CRISPR/Cas9 blocking...DNA region? To this end we developed a technology called CORRECT, which, by undergoing two rounds of gene-editing...efficiently incorporated during HDR. We decided to systematically characterize this phenomenon and found a general...two successful mutations in a single cell. We calculated the likelihood of obtaining homo- and heterozygous...University. He currently works to understand pathological mechanisms of Alzheimer’s disease by developing...

...transformative biological discoveries and life-saving medical breakthroughs and undermine critical research ...Addgene as a trusted partner for archiving their biological materials during funding transitions. Enabling...private partners to create additional pathways for critical scientific work to continue.  Moving Forward ...

...-orbital AAV injection at 4 weeks. Cortical layers (L1-6) and scale bar are indicated. B) Specificity,... core, 2–4 glycosaminoglycans (GAG), and are (typically) tethered to the cell surface by a GPI-anchor.... types in the brain can be a challenging — but critical — factor for successful experiments or therapies...enhancer-AAV vectors targeting specific classes of cortical and striatal interneurons (Furlanis, Dai, Leyva...PV-positive cells out of all BiPVe3-dTom cells. C) Confocal stack of a biocytin-filled cell expressing BiPVe3...

...one be interested in state machine technology? Biological research has produced a massive amount of information...processes that are influenced by temporally ordered biochemical events. How do RSMs work? The state of an RSM...prescribed region of DNA termed the “register”. Each chemical input to an RSM drives expression of a distinct...to record all permuted substrings of a set of chemicals. Specific examples of arrangements that should... record all permuted substrings of up to seven chemical inputs are given in Table S2 of our paper. The... He is broadly interested in biocomputing and biological circuits.     References 1. N. Roquet et al.,...

...Translocation Reporters (KTRs) whose cellular localization serves as a proxy measurement of kinase activity...KTRs) Kinase Translocation Reporters (KTRs) are genetically encoded fluorescent kinase reporters. A KTR is...which is further fused to a bipartite nuclear localization signal (bNLS) and a nuclear export signal (NES...phosphorylatable. Prior to phosphorylation, the KTR localizes to the nucleus, but following phosphorylation ...breakdown (without the nucleus, there’s no nuclear localization to monitor). Whether you’re looking for a ready-to-use...

... first, which he called aequorin, gives off a faint blue light when it binds calcium ions, and the second...second, which we now call GFP, absorbs that blue light and glows green. In the late 1980s, another researcher...gene transcription, as well as to track protein localization. And as it turns out, he was right! He began.... He even obtained samples of certain ones from local pet shop aquariums in Moscow where he lived! His...

...re tired of calculating dilutions or you can’t quite remember the hazards of that chemical in your obscure...surpassed 60,000 views for the month of September! Historically we do better in September than in the summer..., Lianna Swanson, and researcher at Sanofi Pharmaceuticals. This post highlights a rare disease, Fibrodysplasia...can find the rest of our September posts in chronological order here: Four Things I Learned at the 10...

...human speech near the laminar airflow, the human scalp, sneezing, coughing, etc. Also, a constant in-flow...contamination Detection of mycoplasma by naked eyes or optical microscope can be very difficult, so how do you...cells detect mycoplasmas, they trigger a series of chemical reactions and this induces a noticeable color ...contamination, before using the cell lines for assay or analytical purposes. Use 0.1-micron filters as are they ...Fumigate the laminar airflow and the lab facility periodically. The CO2 incubators, used for maintaining cell...cleaned regularly and checked for contamination periodically. For more tips, check out Addgene's video Getting...

...by the use of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD...these complexes are so robustly expressed that anatomical assessments can be easily performed too. While...identified this feature of AAVs as a new tool for neuroanatomical tracing (11). With the ability to produce neuron...Retrograde AAV Preps at Addgene 2) Comparison of genetically engineered receptors (i.e. chemogenetics vs. ...should strengthen its appeal for the scientific and medical communities. Many thanks to our guest blogger ... Floris G., et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring...

...depositors; Gizela, the lab member who knows how the physical kits are made and shipped; Chiara, the tech transfer...repository, performing quality control, and providing technical customer service. Team members are highly trained...problem? Test a hypothesis? Whether you officially call it a “project team” or not, most scientific collaborations...But this is a blog post, so here are just a few critical features based on my own experience: Leadership...should be on the team. As a team leader, I’ve (diplomatically) booted people off teams. I’ve also kicked ...like meetings, but run well, meetings can be a critical part of any successful team. For reasons I don...

... developed an improved voltage indicator (GEVI) called JEDI-2P. Using a custom, high-throughput screening... the protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is ...approach and outcomes of the screen for improved genetically-encoded voltage indicators (GEVIs) based on the...optogenetic silencer in experiments with mouse cortical neurons, far out-performing previous attempts ...selective for K+ over other Na+ and other cations. B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer...

...The biomedical field is often concerned with understanding the cause of diseases and how to treat those...optimization is performed to take the drug to the pre-clinical and human trial phases.   So how does CRISPR factor...enables researchers to select and generate physiologically relevant models. In complex diseases, it can...standard is to test the drug on models which are genetically proficient and deficient for the target. CRISPR...discovery – it’s an exciting time to be in the biomedical research field! References and Resources References...

...published data by biomedical researchers and ready access to stably archived large-scale datasets by computational...new freely available resource developed by the Biological General Repository for Interaction Datasets (...CRISPR screen results available to the entire biomedical research community, but also allows cross-interrogation...relevant to screen result interpretation. The analytical method for score assignment and significance ...

...antibodies in your lab. Perhaps you’re even in that mythical place where antibodies are well-organized, documented... sample prep. This involves using heat and/or chemicals to break some of the bonds keeping the protein...epitope (right). Created withBioRender.com.   Biological Processes  If you’re feeling relieved that you...the available epitopes could be affected by, or critical to, the assay.   Next, look for antibodies that... also working for a Western because they both typically use denatured protein samples. If, however, you...

...reproducibility and an inability to improve upon these critical research tools. Addgene is excited to announce...reagent resource for the neuroscience community called NABOR (Neuroscience AntiBody Open Resource). We...partnered with Dr. James Trimmer of the University of California Davis School of Medicine on this grant. Dr. Trimmer...in neuroscience research today, but there is a critical unmet need for an open-access, molecularly defined...such as plasmids and viral vectors. However, it typically takes a few years for a new project to reach a...