...CRISPR screens check out Addgene’s 2016 publication “Practical Considerations for Using Pooled Lentiviral...experiments you use them for! A well-developed biological question and experimental system are absolutely...coli, and T. gondii genes.  2) What genetic modification are you trying to make? Addgene carries CRISPR...next-generation sequencing to check the quality of your amplification. If you've obtained a ready-to-use lentiviral... As such, sequencing CRISPR libraries after amplification and after a screen requires the use of next-... to skewed results. Selecting a cell type - Theoretically, any cell type can be used in a CRISPR screen...experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen your results...

...When Jessica Sacher, a microbiologist from the University of Alberta, saw that scientists were using ... request for phages to the directory does not typically come out until nearly the last minute when all...all other options have been exhausted. Theoretically, phage alerts can be sent out to the directory earlier...team must decide which labs should receive the clinical sample to test their phages. Because the sample...phages targeting organisms closely related to the clinical sample. For treatment, it is best to identify ...researcher may only know how to prepare phage for one application but not another or they may have only used phage...phage in basic research but not therapeutically: not all labs in Phage Directory have the resources or...

...Regulatory DNA function is typically inferred based on correlative biochemical characteristics and ectopic...Canver of Boston Children’s Hospital and Harvard Medical School. Addgene is proud to present a video reprint... of Visualized Experiments (JOVE). The video publication by Stuart Orkin and Daniel Bauer's labs details... size and frequency [3]. There are several applications in which the production of deletions by paired...useful to study haploinsufficiency. Subsequent modifications at the remaining allele within the deletion ...for deletion screening by conventional PCR. Biallelically disrupted cells can be easily identified. Even...investigator at Boston Children’s Hospital and Harvard Medical School. His lab studies the molecular genetics ...

...my opinion, it was more important to communicate the applications of CRISPR over the details of how it...m a father of three, a scientist and science communicator, and I strive to be a volunteer and contribute...parents who can answer some of the surprisingly technical questions elementary students can ask. It doesn...unlike elementary schools, middle schools have dedicated science teachers. I tried contacting our school...simplistic and I would insult the students. Too complicated and I would quickly lose their interest. I won...great feedback from teachers and other science communicators.  What can CRISPR do? I spent more time on... dangers of people using the technology in an unethical manner, but I also tried not to be an alarmist...

...fixatives - chemical cross-linkers and organic solvents.  Chemical Cross-Linkers Chemical cross-linkers...of Fixatives in Histology. Leica Biosystems. https://www.leicabiosystems.com/us/knowledge-pathway/fixation-and-fixatives...-linking cellular proteins. Cross-linking is a chemical reaction that covalently joins two molecules and...proteins can be lost during fixation with these chemicals, which impacts sample structure and could also... target to be washed away. Furthermore, these chemicals can also alter the structure of proteins, which...Tween-20, or saponin. Each detergent has different chemical properties that will impact the level of permeabilization... don’t even involve antibodies, they are still critical to the outcome of your IF experiment and deserve...

...scientists are overrepresented on Twitter Mathematical, life, and physical scientists are underrepresented. It...mostly because of this article: “A systematic identification and analysis of scientists on Twitter.” This...from the United States Standard Occupational Classification System  and Wikipedia. 2. Social and computer...Nature, Sciencemag, Wiley, Cell and Scientific American Figure 4: Top Scientific Domains Tweeted by Biologists...shown. Top: The follower network of life (green), physical (blue) and social (orange)  scientists largely...disciplines: social (left), life (center), and physical (right). How do you use Twitter as a scientist... Y., & Sugimoto, C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos...

...viral vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation of specific...of these discussions as a publicly accessible practical guide which was recently published in Frontiers...of molecular tools and sensors Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT...transgene expression by restricting expression to genetically defined cell types. Specific transgene expression...defined set of neurons can then be targeted with optical stimuli. The restricted light exposure to a specific...result in opsin expression in their cell bodies. Optical stimulation in a different brain area can then ... more versatile. To complement the wealth of genetically encoded tools available, researchers are also...

...for both transient and stable gene expression applications in plants. This smaller, easily customizable...Agrobacterium are termed binary vectors because they must replicate in two species: Agrobacterium and E. coli. Like.... coli. However, they also need to be able to replicate in Agrobacterium to ensure that they are maintained... Thus, binary vectors contain two origins of replication, one each for E. coli and Agrobacterium, along...bases each. By switching the E. coli origin of replication and codon-optimizing the Agrobacterium ORI, they...constitutively active M24 promoter has been modified with duplicated enhancer domains and characterized in studies...these properties of pSiM24 are suitable for many applications, the plasmid can also be customized due to its...

...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...

...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...

... 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...

...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 ...

... 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...

...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...