...guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc...professional development I had a wonderfully supportive mentor and graduate office. I cannot stress how important...my blog helped immensely when CINSO started brainstorming the resources we wanted to provide online. ...in real time! I follow great online science communicators like Phil Plait @BadAstronomer, Emily Graslie...when I was trying to find local informal science educators, and ever since, the State Museum has been the...week. I gained experience explaining science to visitors of all ages, learned a lot about the dinosaurs...Melanie Fox is currently the founder and executive director of Central Indiana Science Outreach and a Postdoc...

...making it easier to construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed ...cloned into a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector...just upstream of a stop codon. Briefly, the PITCh vector should be constructed with 5’ and 3’ microhomology...PITCh Step 1: Generate microhomologies in the PITCh vector ~20 bp 5’ and 3’ microhomologies are added to ..., and this construct is inserted into the PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an... vector carrying Cas9 and the PITCh- and locus-specific gRNAs Step 4: Select for puromycin resistant cells...

...conjunction with activator or repressor domains to control gene expression. Base editors can help you make... in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene (e.g. ...experiment As with any experiment, there are many factors that need to be considered during the planning ... that would work for your system. Addgene has depositor submitted protocols and links to a CRISPR forum...Components of System Application Mammalian expression vector Promoter driving Cas9 expression can be constitutive...CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles. Transient or...

...Science describing the development of “transcriptors,” transistor-like digital genetic switches that enable...Stanford develops engineered DNA systems capable of storing data and computing inside living cells. He's also...cellular computing. These logic gates built from transcriptors are now available at Addgene through the Biobrick...world using words like OR and AND. These logical operators are so useful they are implemented in most materials...the beginnings of computational logic and data storage. What we don't have are the keyboards and displays...mining. There is an absolutely incredible hidden story here. The parts we are releasing simply plug into... plants. We are really plugging into the "stuff sector" if you will, making chemicals and materials, and...

...signaling pathway with many receptors, signaling molecules, transcription factors, and ligands. The presenter... unbiased discovery. With these screens, the investigator generates a cell population where all genes ...contexts. Most of the immune cells we study are migratory, and their exact localization is important for...types of interactions to a large extent involve receptors expressed on the cell surface. Thus, a screen ...targets, preferentially those for which validated inhibitors or even approved drugs already exist. If you ...The gRNA oligos are subsequently cloned into your vector of choice using the adapter sequences that you ...strategies related to other nucleases as well as inhibitory and activating approaches.  Many thanks to our...

... for depositing labs to its website, allowing depositors to download their plasmid request data, as a ...information can be of personal interest to the depositor, but it can also be valuable data to share in ...total requests, total countries represented by requestors, year of first reagent sent, year with most reagents...is launched, it will show my data by default. Depositors can download their CSV file from Addgene by following...Dashboard After the upload, the name of the depositor and the date of downloading the CSV is shown (...and the source code is available in the Github repository (you can find the link in the upper right hand...dashboard useful when analyzing and sharing your own depositor metrics!  About the Author Joachim Goedhart is...

...including the internet!) can vary depending on factors like socioeconomic status and geographic location...communication should be fun and engaging for the communicator as well as the audience, so pick a mode of communication...restrictions (or both!) will be a major limiting factor at multiple points. Before you embark on a project... convey enough information to tell a compelling story in an image or do you need a full exhibit? In a ....  Static. Can be difficult to convey an entire story or context via still images. Some things cannot ...information. Primarily useful for data-focused stories. In-person lectures, presentations, etc… Low... Low barrier to entry for both audience and communicators. Well-developed content creation platforms.  ...

...What do a viral vector production facility, food allergy testing lab, and the grad student down the hall... curves in their day-to-day work. Indeed, viral vector production facilities frequently use qPCR with ...with each step of the series diluted by the same factor. For example, Figure 1 depicts a 2-fold dilution... standard curves vary depending on a number of factors, including the user, equipment, assay incubation...measure.  Dynamic range depends on a number of factors, including the dilution series of the standard ...standard curve dilution series with lower dilution factors. If the unknown sample’s measurement is too high...assays you will be using ahead of time to see what factors may impact the dynamic range of the assay. Try ...

... the notation works? The + operator denotes intersection and the – operator denotes set difference. That...inclusive markers of regulatory regions in the genome, including enhancers, promoters, insulators, and more. I...proteins”) for NANOG, an essential transcription factor in embryonic stem cells (ESCs). Your ChIP-seq finds...of embryonic stem cells (ESCs) with a lentiviral vector, an attenuated retrovirus in the same family as...enhancers, lncRNAs, repetitive elements, repressors, insulators, and more. We know that this matter exists, but...criteria: no TTTTT allowed (this is a polymerase terminator), and no off-target effects for the identified...

...keeping track of speakers. During your graduate or postdoctoral work there are loads of opportunities to create... concrete happen. You may have more willing collaborators than you realize. Being at a university provides...meetings and take minutes. Get to know your collaborators from other institutions personally on conference...relationship with the perspective of my potential collaborators in mind. Being at a university allows you to...with people at your organization. You can meet collaborators  for lunch, make a point of talking to them ... different countries). Here social media is a connector and virtual meeting place: You can follow interesting...as messages and keep you up-to-date on your collaborators’ professional developments without directly ...

...us. Use a well working vector system This allows you to quickly assemble vectors and test different ideas...MoClo system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo...to quickly screen and investigate transcription factor candidates and the regulation of genes in the biosynthetic... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...and a vacuum is applied with a vacuum bell or desiccator. Successful infiltration can often be seen by...system you can easily and quickly create customized vectors. For more information on MoClo for plants, check...

...-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School... Darrell recently joined the Addgene Board of Directors where he hopes to help keep us focused on our ...Darrell's goals as a member of the Addgene board of directors - increasing open source sharing and helping us...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...

...they do so is through G protein-coupled receptors (GPCRs), receptors that play many roles in signaling processes...which a donor fluorophore is used to excite an acceptor fluorophore. The difference for BRET is that the... between a luminescent donor and a fluorescent acceptor. TRUPATH uses an updated version of BRET called...constructing 14 luciferase donor Rluc8-Gα chimeras, four acceptor GFP2-Gγ chimeras, and two untagged Gβ constructs...non-uniform across the transducerome for a specific receptor-ligand pair. They further used TRUPATH to analyze...Mitaku S (2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters...

... referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may ...plasmid with its elements described below.   Vector Element Description Origin of Replication (ORI...the target gene. Vital component for expression vectors: determines which cell types the gene is expressed...Regardless, once the cloning steps are complete, the vector containing the newly inserted gene is transformed...Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of a chemical in a particular environment...about molecular cloning, how to choose a plasmid vector, molecular biology tools and references, and how...

...upon an indicator they developed and published in 2010 for visualizing the transcription factor cAMP-responsive...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since the...Griesbeck is motivated by a particular interest in monitoring the activity and biochemistry of living neurons...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...while in principle adapting a FRET sensor for monitoring calcium versus zinc should require the simple... variants of mOrange2 and mCherry as donor and acceptor domains. He says, these new colors might now enable...

..., the researchers did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform... study has just been uploaded to the preprint repository BioRxiv. This example clearly illustrates the...  Figure 2: When a researcher needs a collaborator for a specific task, Crowdfight will: 1) Find...researchers, such as postdocs and principal investigators. They also come from many different scientific...website. Once curated by our in-house team of collaborators, the requests were forwarded to our volunteer...IgnacioAmigoH     María Hernández-Sánchez is a postdoctoral researcher at Instituto de Investigación Biomédica...Biomédica de Salamanca (IBSAL) and one of the coordinators at Crowdfight. She is currently based in Spain...

...Scientific Illustrator Plant Breeder Zoological Worker Conservationist Documentary Director Parasitology...skills include: Science Reporter Scientific Illustrator Research and Development Scientific Advisor...Lawyer Technical Writer Meteorologist Journal Editor Textile Technologist/Designer     Fieldwork...Bioinformatician Science Writer Grant Writer Journal Editor Science Marketer Software Developer Learn Science..., and how many jobs are normally available (as sectors such as forensics currently have too many applicants...hobbies and volunteer experiences, as well as work history can really show that you have some of the abilities...

...This will be highly dependent on vector dose. The color diversity in Vector Assisted Spectral Tracing (VAST...catch our April AAV webinar with Tim Miles, PhD, Director of the CLOVER Center at CalTech? If so, you may...drop in transduction coverage compared to just one vector.   Q2: Do you think it is possible to make an efficient...this scenario, you may be best served by a dual vector, split intein approach.   Q3: What’s the best method...concentrated to ~1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011...Plasmids Adenovirus Guide Page Subscribe to Viral Vectors Blog Posts ...

...Oleispira antarctica, and the P. haloplanktis Trigger Factor), that would allow the stable folding and expression...compared to the E. coli strain expressing the empty vector. And thus was born ELSA - our happy lil’ cold-tolerant...of stakeholders from industrial and commercial operators, government agencies, active researchers, as well...interact with personnel from the industry and brainstorm with each other. The event also featured a seminar...The talk highlighted the importance of reagent repositories and the myriad facilities offered by them which...understanding of the societal ramifications of laboratory science, and how it may best be leveraged for...re always delighted to engage with potential collaborators and interested audiences. Please visit our team...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...regulate botulinum neurotoxin were also created. The depositors demonstrated the utility of light-mediated suppression...slices from mouse, and showed that it can alter motor responses in C. elegans. Find the split BoTN/B plasmids...acoustic reporter gene (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These.... 2018. PubMed PMID: 29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated ...microRNAs (miRNAs) are recognized as important regulators of post-transcriptional gene expression across...They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert site and a reporter...