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...controlled by collections of neurons that are intricately wired into circuits through synaptic connections...connections to that population. Limitations of common genetically encoded neuronal tracers Neuronal tracers such...neurons. As compared to epitopes tags, FPs are intrinsically fluorescent and can therefore be used in live.... Indeed, to create the smFPs Looger’s team strategically inserted 10 to 15 copies of single epitope tags...darkened chromophores. These probes are both intrinsically fluorescent and can be detected with highly ...photostable than GFP. In addition, antibody-based amplification of their signals is weak and increases background...postsynaptic cell adhesion protein that plays a critical role in neural development. It belongs to a huge...

...range of logistical barriers. Addgene worked to identify, reduce, and remove these logistical barriers...’ve turned the available into the accessible. Physical access Addgene is located in the USA, but our ...Beijing Zhongyuan 2012 China Summit Pharmaceuticals International 2013 Japan Leehyo Bioscience... items to countries with a distributor. Arrows indicate the year we entered into an agreement with a distributor...Systems AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your gRNA for...Digest Analysis We also introduced guides — practical primers to help users get started with tools like... country are represented in blue; darker blue indicates more views from that country.   While written ...

... approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger...efficient for short, unstructured templates and significantly reduced undesired edits (indels, etc.), while...Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...delivery by modular envelope design. Nature Communications, 14(1), 5141. https://doi.org/10.1038/s41467...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting...control over when and where the enzyme is active, drastically reducing background labeling caused by the biotin...

... well as neural development. Specifically, SHH is a morphogen, a chemical signal that diffuses down a ...exhibit abnormal segmentation during embryogenesis. Typically, regularly spaced denticles (bristles) outline...on the spindle of a spinning wheel but also a biological transposon system. Sleeping Beauty is a synthetic...transposon system including those from the original publication, Ivics et al 1997. Spaghetti Squash, a regulator...contract depending on the context. Contraction is critical for processes such as cell movement/motility, ...bunch of cookie monster eyes.”  Check out the publication to see for yourself. We have one plasmid in our...embryo (Azplazu and Frasch, 1993). Tinman is specifically involved in the formation of the heart amongst...

...Young is a Medical Writer with a PhD in reproductive biology and genetics. She loves to communicate the latest...method for selecting the guide RNA, validating its efficacy in vitro, and using it in mouse embryos to generate...control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted from Mashiko...plasmid using standard cloning techniques (The XX indicates where your target is inserted, disrupting the ...plasmid, see Mashiko et al., 2013). Testing the efficacy of your gRNAs is then a simple matter of transfecting...effective ones to increase your chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs...et al., 2014). The plasmid is sturdier than the delicate RNA and therefore there has less chance of contamination...

...graduate students and postdocs to think analytically and critically. Passion fuels the drive to continue ... Connie Cepko (Harvard Medical School) , and Dr. George Church (Harvard Medical School) for taking the...people (especially you!) care about. Have you critically evaluated your plan? Have you read the relevant...write a grant for this project, what would be your critical experiments and why? What techniques and skills...help move your project forward? Most projects typically require a diverse set of skills. It is important...direction of the project? What is your timeline? A typical research project can take several years to complete...time to generate new ideas and think about them critically before diving in. Research projects are designed...

...transcriptase (RT), MarathonRT. Reverse transcriptases are critical tools for many fields of biology and enable the...MarathonRT, isolated from Eubacterium rectale, significantly outperforms other RTs in this regard and is ...Nanobodies are single domain antibody fragments typically engineered from heavy-chain only antibodies found...makes them more adaptable to a broad range of applications. Recently, the Spiess lab developed a set of... with specific genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered...isolated and analysed. In this work, the authors specifically investigated protein binding at the TERT gene...his group from the University of Massachusetts Medical School have developed a group of plasmids (here...

..., soon directed himself to finding “immunological and virological solutions to crises like those and ended...Therapeutics, which was acquired by Ultragenyx Pharmaceutical in 2017. As one of the original employees at...learned more about Addgene. “The mission to share critical reagents such as plasmids and viral stocks for...depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function of all 20,000 genes in disease...Target Discovery at the Novartis Institutes for Biomedical Research. Drug development was something Stegmeier...genetics.” As a postdoc in Stephen Elledge at Harvard Medical School, he developed the first lentiviral microRNA-based... interference. In the first two months since publication, the lab got over 100 requests for the plasmids...

...  After identification of ABE activity, XMAS-TREE can also be used to purify these modifications and work...system by using a stronger promoter and different biochemical secondary energy sources to increase protein ...DNA of interest, and cell extract.    Graphical Abstract: Schematic representation of cell-free...ABE-targeted cells By Ellie Adams Single base modifications through ABE or CBE's have the potential ability...point mutations. They allow single-nucleotide modifications to edit DNA without requiring a double-stranded...difficult to modify in the past.   Figure 1: Identification and enrichment use of XMAS-TREE. a) Cells are...mCherry and GFP expression to check for ABE’s. b) Quantification of base-editing efficiency using XMAS-TREE-...

...1952, Joshua Lederberg set out to clarify the classification of these cytoplasmic inheritance factors. He...hereditary determinant”. His proposal, however, was basically ignored. A separate term, “episome”, defined as... plasmids have greatly impacted many areas of biological research and have been key in advancing our knowledge... as bacterial conjugation and recombination, replication and topology, and cloning and gene expression...National Academy of Sciences of the United States of America 110.39 (2013): 15521–15529. PMC. PubMed PMID: 24043817...Joshua. "Cell Genetics and Hereditary Symbiosis" Physiological Reviews 32.4 (1952) 403-430. Pubmed PMID: 13003535...PMC2134509. Cohen, Stanley N. et al. “Construction of Biologically Functional Bacterial Plasmids In Vitro.” Proceedings...

...for Neurobiology has been working on genetically encoded indicators of calcium and other small molecules...of interest,” Griesbeck said. Because they are chemical compounds, they have to be applied or injected...lymphocytes, too, making them a versatile tool for application in both neuroscience and immunology. Don’t be...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...give it a shot. “Beginners sometimes think it’s complicated, but actually it is very good,” he said. “It ...his team is now applying it to improve upon an indicator they developed and published in 2010 for visualizing...tools useful, including his zinc sensor and his genetically encoded magnesium sensor, the first of its kind...

...humanistic and philosophical implications. As an example, a group at Caltech used an optical fiber together...handle on some of these mysteries. Another application is clinical; can we find ways to treat patients? Over...Optogenetics is a toolset that lets you turn the electrical activity of neurons on or off using pulses of...photosynthetic or photosensory molecules that come from basically every branch on the tree of life. Our most recent...implant already. If we can use light to more specifically activate a set of neurons, maybe we could treat...Addgene Alerts box. References: Independent Optical Excitation of Distinct Neural Populations. Nathan...

... regions of homology at their ends, which are typically created by PCR. Then, the fragments are incubated...neighboring fragments can be created via PCR amplification with primers that contain the appropriate homologous... tools that help you design primers for PCR amplification of fragments to incorporate such regions of ...competent cells. The success rate of this reaction is typically fairly high, so there is usually no need to screen...assembly meets CRISPR Gibson can be adapted to more complicated cloning schemes, such as those where the vector...CRISPR technology and is described in a recent publication by the Lockey lab (Wang, et al. 2015). In this...the bacterial chromosome (up to 100 kb) were specifically cut out via CRISPR and then assembled into a...

...improve our shipment processes. After reading a publication describing how short term storage on dry ice ...aqueous samples might be affected by dry ice, and specifically how dry ice can affect virus from Addgene. ...medium, which is buffered and contains the pH indicator phenol red. We froze samples in a -80°C freezer...closed dry ice storage container for 48 hours (to replicate how our viruses are shipped), where they would...exposed to CO2/dry ice. All samples were tested in duplicate. What we found Not all tubes (or rather, not ... the samples were at the control pH of 7.4 as indicated by color (data not shown). What we did next: Venting... at -80°C. We then thawed the tubes after the indicated incubation periods, and measured the pH. We found...

...Talley Lambert, a Research Associate at Harvard Medical School. The need for a community fluorescent protein...one of the most indispensable tools in modern biological research.  Any microscopist will tell you that...non-trivial, however, as FPs are tremendously complicated entities with a large range of characteristics...maturation, oligomerization), many of which are dramatically affected by environmental conditions (such as...decision remains spread across literature in publications that introduce these tools. Years ago, Kurt...incorporate FPbase data in their own projects and applications, a REST API allows programmatic retrieval of...Talley Lambert is a Research Associate at Harvard Medical School. He manages the Cell Biology Microscopy ...

...transcription, or DNA repair, or replication, or any number of CRISPR applications and thought “OK, but how does...pages containing lists of plasmids from the indicated publications. You can request all of these plasmids ...to think about DNA in this context, but also specifically study it. They might be researching enhancer...--directly or indirectly--many other areas of biological research. If you’re looking for resources that...you study how chromatin effects your favorite biological phenomenon, there’s a reasonable good chance ...below pull up lists of plasmids containing the indicated gene, and you can subscribe to these pages to ...RANbodies) as versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR Proc Natl ...

...proof-of-concept for pre-clinical and clinical applications of dCasRx. The identification of Type VI Cas13d ...RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader...even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable...efficiency; where shRNA treatment produced 500-900 significant off-targets, CasRx displayed zero. Unlike Cas9...target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding properties...enhancers. After AAV-mediated delivery to iPS-derived cortical neurons, dCasRx-mediated exon skipping improved...

...comparable in efficacy to dCas12a systems in both gene activation and base editing applications.  Figure...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...members or students new to protein expression and purification. Figure 3: The Penn State Ladder run ...challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature CRISPR-Cas system ...: Overview of CasMINI for genome engineering applications. Image from Xiaoshu, et. al. 2021.   Created...

...population (typically cells or small organisms) and then assaying for a functional or phenotypical outcome... daunting task of finding new factors in your biological system of interest, the so-called “fishing expedition...that both attempt to identify new players in a biological process. Players can be proteins, genes, enzymes...obligatory growth advantage – that is, you are not specifically looking at cells that can survive. Instead, ..., the entirety of the small molecule realm is typically not feasible to test in any given screening experiment...selection process. Non-functional traits will be automatically eliminated, unlike in screens. Selections begin...the answer that much faster? In reality, many biological questions are not conducive to selections and...

...their idea is. While communication styles in academia and industry vary drastically, he reassures that ... As a biomedical engineer and scientist, I like to envision that my research will have a direct impact...competitions. After much learning, the focus of the application was changed, but Sample6 still leverages the ...fit for this set of technologies, effectively communicating the advantages embodied, and driving the enterprise...He explained that entrepreneurs must clearly communicate their idea at various levels of complexity and...biggest challenge during this time was learning to communicate effectively to non-scientific audiences. "There...for the company; this will help avoid future complications and potential conflicts that waste money and...