...C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use ...insert will typically contain both a protein modification sequence and a selectable gene product for isolation...tags have been integrated. Simply design your amplification primers with the desired targeting homology—.... cerevisiae, looking at their performance in categories such as brightness, stability, and disruption...optimal FP combinations for use in yeast imaging, categorized by specific filter sets and experimental output...This collection is ideal for tandem affinity purification of protein complexes. John Pringle and Jürg ...deposited a large collection of plasmids for genome modification in yeast developed by Dr. Pringle’s former lab...

...this paper at Addgene. Other relevant blog posts 3 Challenges in Plant Synthetic Biology Quick, Versatile...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...

...regions of dsDNA. To generate a 3’ hydroxyl group needed for DNA replication, Rep78/68 binds the Rep binding...Rep78/68 play a key role in AAV genome replication. Replication initiates from the AAV genome ITRs on ...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid (cap) gene. In wild-type...its products. Created with BioRender.com   Replication (rep) gene The rep gene encodes four proteins...proteins that are required for viral genome replication and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene...nicked by its endonuclease domain. Following replication, Rep78/68 stays attached to the AAV genome and...

... ways we get your plasmids out there Beyond these 3 big pushes, we try to increase your work’s visibility...Addgene ID number, your publication will be listed on our website as a “Publication Citing this Plasmid” ...upon other researchers being able to find their publications in the first place. As a curator of the thousands...make it easier for you to find the plasmids and publications relevant to your research interests. We do this... this by promoting citation, supporting sophisticated search, and curating plasmid collections. Addgene's...number as well as the reference for its original publication. To help facilitate this process, we explain ...fruits of your labor and won’t have to repeat the modification. This will save both time and money. A quick...

...interest.   Figure 3: Each publication featured in BioGRID ORCS has a dedicated summary page with PubMed...favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section...manually curated and annotated from more than 45 publications, beginning with the seminal research articles...screen curated in ORCS is linked to the original publication via its PubMed identifier, which can also be ...a list of individual screens curated for the publication. Clicking on the screen name takes users to a... supplemental data tables available with the publication are also provided for the user. This approach...

...Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM, Patterson...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...

...fluorescent proteins. Krumholz et al (Sci Rep. 2014 Feb 3; 4:3939. doi: 10.1038/srep03939.)  ...of hues. While those tools are great for many applications, Verkhusha and his lab at Albert Einstein College...light. As the researchers described in Nature Communications last year, “The capability to control spectral...Verkhusha’s team reported another advance in the application of near-infrared fluorescent proteins, iRFP670...fluorescence is because the light becomes very scattered,” Verkhusha explained. Ultrasound wavelengths,...hand, are larger than the size of a cell, so they scatter much less. As a result, ultrasound enables higher...

...plasmids are categorized into a 3-level system on our website:   A yellow flame indicates that a plasmid...re proud to honor the creativity, passion, and dedication to sharing epitomized by the creators of these...Since we so sincerely appreciate our depositor’s dedication to scientific sharing, we wanted to celebrate...

...system for signal amplification in gene expression and fluorescence imaging.” Cell 159(3) (2014):635-46....aggregation, it’s recommended for most imaging applications. What can you do with SunTag? Tanenbaum et al...activation of target genes, but we’ll save that application for another day! References 1. Tanenbaum, Marvin...

...with others in the IBEX community.   Figure 3: Screenshot showing recent topics in the IBEX discussion...something doesn’t work, it’s not a failure. It’s an education.   Figure 4: Zenodo page showing the IBEX... repository for reagents, protocols, panels, publications, software, and datasets. It’s also a great place...

... 6–8 weeks Average shipping time: 1–2 days (USA); 3–4 days (international; varies widely by country and...request is approved, you will receive an email notification that your request is ready to order. An “Add...subscribe to the Addgene blog for updates and educational resources.  We hope you’re as excited about this...

...it doesn’t help much if the actual response takes 3 days and still doesn’t even answer your question. ...right ones, quickly, and this is why we have a dedicated support team that can help you through any search...webpage, you can browse through our various FAQ categories or search through the entire FAQ collection. ...

... cells, as well as in cortical neurons in layer 2/3 of the mouse brain. Rubin’s work shows that the yeast...Gerald Rubin’s lab sought to make complex genome modifications in Drosophila using multiple recombinases. To...offspring carried both the recombinase and GAL4, indicating that the recombinases were inherited normally...

...project. Furthermore, Group Leader position applications require 3 letters of reference- the graduate school...yourself to and talk with more senior scientists. 3. Work intensely, but intelligently Before you do your...need? Multiple first or corresponding author publications (published, not in preparation) A history of...The other items will follow. If you get more publications, you will be more competitive for fellowships...to write as well as mentor. Yet, even the most dedicated mentor is not always available for mentees due...interests, complementary perspectives, a shared communication style. You “get” each other. The other factor...makes time for you, as needed, this is a strong indicator that this person cares about your science and ...

... term sgRNA. Seed sequence: The 8-10 bases on the 3’ end of an RNA guide (either crRNA or sgRNA). Cas9...simplifying the CRISPR-Cas9 system for laboratory applications. At Addgene, we sometimes refer to the programmable...

...signaling pathway is unknown.      Figure 3: Chimeric OptoXR incorporating rhodopsin undergoes...Ultrafast optogenetic control. Nature Neuroscience, 13(3), 387–392. https://doi.org/10.1038/nn.2495 Marshel...channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from the archaebacteria Chlamydomonas ...stimulation. GPCR signaling GPCR signaling is a complicated process involving many downstream signaling mechanisms...break your experiments!  First, closely read the publication, including the supplemental data. You’re most... both of which have extensive collections of publications and information on hardware, optogenetics tools...2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of the ...

...Fragment Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies...molecule chaperone Skp. Front Cell Infect Microbiol 3: . https://doi.org/10.3389/fcimb.2013.00072 Additional...tool that can be used for a variety of research applications. Within the immune system, antibodies are produced...antibodies to bind proteins is useful for research applications as well because they allow scientists to target...150 kDa, a nanobody is 12-15 kDa.   Research applications for antibodies You can use antibodies for experiments...Let’s dive in and take a look at some of these applications. Western blot Western blots are used to detect... by binding to cell surface receptors. These applications are typically used for investigating signaling...

...while AAV-PHP.S is typically used at a dose between 3 x 1011 and 1 x 1012 vg*. Most systemic capsids are...transduction in New World monkeys   Figure 3: Evolutionary tree depicting the phylogenetic relationship...414 (2019). https://doi.org/10.1038/s41596-018-0097-3 More resources on the Addgene blog Viral Vectors 101...Sensory Ganglion Neurons Using AAV-PHP.S. eNeuro,9(3), ENEURO.0373-21.2022. https://doi.org/10.1523/ENEURO...Are Limited to C57BL/6J Mice. Molecular Therapy, 26(3), 664–668. https://doi.org/10.1016/j.ymthe.2018.01.018... 2023 * indicates viral preps available. Mechanisms of crossing the BBB are indicated in parentheses...suitability of a systemic capsid for an individual application is dependent on a number of factors, including...

... research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini (6), and ...PMID: 26780180. PubMed Central PMCID: PMC4744125. 3. Sanjana, Neville E., Ophir Shalem, and Feng Zhang...the user. This approach is useful for several applications as will be discussed below. The software could...contain multiple gRNAs targeting all genes in the indicated species and were suggested to be the best gRNAs...

...10.1007/978-3-319-07758-1_2 More resources on the Addgene blog Viral Vectors 101: Viral Applications Viral ...methods of replication. In fact, an entire classification scheme, known as the Baltimore replication classes...that in a bit! Viral genomes and replication When it comes to replication, viruses tend to march to the ...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit: ...an intermediary DNA replication step between DNA and mRNA. DNA genome replication can involve an RNA or...cell. The cell then uses its own machinery to replicate the viral genome and proteins. This is, for the...before being translated into proteins. RNA viruses replicate their genomes using a variety of RNA or DNA intermediaries...