...microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The...microenvironment’s physiology is deeply connected to these 3 stages and contains multiple cell types: fibroblasts... these stromal cell modifications, stromal cells are also constantly communicating with a tumor, proving... was contributed by Bárbara Pinho, a science communicator at the Portuguese science museum "Fábrica Centro...Proficient in transportation, exosomes are tools of communication and provide malignancy to the microenvironment... Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science museum "Fábrica Centro...

...help ensure ensure equal distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep...With barcode tracking, you will always be able to locate materials quickly and easily. Learn more about ...Additional Resources on the Addgene Blog New Educational Resource: The Addgene Videos Page 5 Science Rap...

...an improved design of the guide RNA that caps the 3’ end with a structured RNA motif. These engineered...editing tools, the desired edits are templated by the 3′ extension of the pegRNAs. PEDAR is capable of correcting...AsCas12f1. They found that this system, without any modifications, is effective in genome editing in both bacteria...therapeutically relevant gene targets and expand the application to the possible treatment of genetic disease....

... motivated reasoning. Psychological bulletin, 108(3), 480. More resources on the Addgene blog Summer SciComm...Further, when science is communicated in a format that is accessible, that communication often neglects the ...SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication: Science Writing... to save a few days’ worth of capsules to self-medicate with when they get a cold. A primary school teacher...dihydrogen monoxide (H2O). Studying science communication as a PhD student Fortunately, by the time I ...Otago had established the Centre for Science communication and I was accepted as a grad student. I chose...a phenomenon called “motivated reasoning,” the catchall term for a range of neat little tricks our minds...

...Minimal Tol2 or miniTol2 is a truncated version of the original Tol2 and has ~3-fold increase in transposition...the QCascade DNA-targeting module with the crRNA, 3) a transposase complex made up by three transposase... up to 10 kB in size in bacteria.   Figure 3: INTEGRATE system for RNA guided transposon insertion... It generally stays put, which makes it easy to locate in a genome. But there is a type of mobile DNA,... hyperactive and likes to jump around from one location in the genome to another. This jumping is what...most transposable elements (TEs) results in the duplication of this target site sequence at the site of insertion...This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme...

...GpNLuc and OgNLuc are available through Addgene (Cat.# 70185 and 70186, respectively). Antares Chu, J...construct for Antares is available through Addgene (Cat. # 74279). Enhanced Nano-Lanterns  Suzuki, K., et...wavelength above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc...PMID: 27465542. PubMed Central PMCID: PMC4974478. 3. Schaub, F.X., et al. (2015) Fluorophore-NanoLuc BRET...et al (2). In this post, I’ll cover two great applications of the NLuc technology. The first couples NLuc...fluorescence. The paper reports a wide variety of applications of the Enhanced Nano-Lanterns including monitoring...2017) Genetically encoded bioluminescent voltage indicator for multi-purpose use in wide range of bioimaging...

...the stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013)...every CRISPR experiment. Off-target events due to >3 bp mismatches or that are sequence-independent are...PMID: 25566793. PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening...off-target sites across the genome and pinpoint the location of mismatches based on the sequences of the genome... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...those relating to DNA structure (e.g. epigenetic modification, bulges) that may also impact off-target edits...cell-based methods are genome-wide, unbiased identification of double-strand breaks (DSBs) evaluated by...

...manuscript review/submission process at JEI. Chapter 3 - JEI target audience and their current readership...Additional Resources from the Addgene Blog Find education resources Use our Plasmids 101 eBook to teach ...

...alterations before submitting - handing them your application 3 days before the deadline isn’t going to be much...work and possibly a new perspective on the field. 3. Highlight your network and your collaborators It’...Series 4. Become a science communicator More and more fellowship applications are asking for a 200-300 ...good quality application. Even following this advice, only about 50% of my grant applications have been ...do need to be able to research and write your application, then give it to someone else to read and / or...it). It’s easy for reviewers to spot when an application is rushed (whether the idea isn’t properly researched...researched / developed or the application is full of typos) and these are unlikely to get past the initial...

...materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various steps ...doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972. 3. Britt JP, et al. Use of channelrhodopsin for activation...There are several important considerations with fabrication of the fiber optics needed for optogenetics. ... for the implant depends on the dorsal/ventral location of your region of stimulation (the z-axis bregma...emitted from the end of the fiber optic implant scatters a great deal once it enters tissue, placement ...0.5mm above the site of your stimulation. The scatter effect is also an important consideration when ...Lasers can be purchased through Laserglow, which caters to optogenetics users and adjusts the lasers appropriately...

...knockout, activation, repression, and barcoding.  3) Are you trying to target every gene in the genome...sequencing. Representation - Most libraries contain 3-6 gRNAs per target gene, and maintaining the distribution... unique compared to the rest of the genome and located immediately 5’ to a protospacer adjacent motif ...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-...experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen your results...

...expression (Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation...work well, and up to 10 base pairs away (especially 3’ of PAM) might work too.  High dCas9 and gRNA expression...approach (spoiler alert: it’s dCas9… with no modifications). Ready to get started?   Figure 1:... when DNMT1 methylates the genome during DNA replication, it doesn’t methylate sites blocked by dCas9....for dCas9 to interfere with DNMT1 even through replication. The dCas9 also needs to be removeable, otherwise...vivo using dCas9–peptide repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol 34, 1060–1065. ...

...plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes...vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require some initial optimization...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct... as resources available, cost and downstream applications. For example, while the PEG precipitation method...

...passion and pay attention to the details. Chapter 3 - Answering interesting biological questions using...Twitter and has helped us update many of our educational resources. Listen to hear all about Professor...speeds science. Chapter 8 - The importance of communicating research findings through social media and other...

... to antibody response 2) very high immunogenicity 3) and long-lasting immune response even after a single...their use in other clinical applications. In the last decade, viral applications in cancer treatment and ...s unpack the myriad number of emerging viral applications and give you some recommendations for putting...putting viruses to work. Viral applications Viruses work by infecting a host cell (the target of viral infection...expressed. It’s important to note that for all the applications we discuss, there is more than one type of virus...retrovirus, AAV, etc.). When you’ve decided on an application, our Viruses as Biological Tools blog can help...viruses and pick one for your experiment. Research applications Gene expression control Researchers are always...

...biological study without her or her family's knowledge. 3. Fossil Rock Anthem One of Tom's earliest and most...advice on how to dive into new forms of science communication. Click here to listen to the interview with ... from Addgene.      Check out more science education tools like Verse Two and Fill in the Planck at...with us. Tom McFadden is a YouTuber and science educator looking for creative ways to implement the next...next generation science standards and communicate science. Learn more about Tom on his youtube channel ...Community Check out All of Our Podcasts Find Great Educational Content in our Plasmids 101 Series Additional...

...degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and ALK) by linking small...with dTAG-13 at doses as high as 20 µM (see Figures 3 and 5 in Nabet et al., Nature Chemical Biology for... and we will provide an update with vendor and catalog numbers. As we are happy to share aliquots of dTAG...

...which is achieved by deleting a large portion of the 3'LTR in the transfer plasmid. The deletion prevents...number of HIV genes required for virus production (3 or 4). Third-generation vectors contain gag, pol, ...lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown... looking for some additional information or clarification on these vectors. Read on to find the answers... thus decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells...type or tissues; this is useful in gene therapy applications where a distinct subset of the cellular population...needs to be targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all)...

...stimulate 2) Fabrication and implantation of the optical fiber into the region of interest 3) Behavioral...thoroughly elsewhere (here and here for instance) [2], [3]. Karl Deisseroth of Stanford University , one of ...10.1146/annurev-neuro-061010-113817. PMID: 21692661.  3. Bernstein JG, Boyden ES. Optogenetic tools for analyzing...Channelrhodopsin2 (Chr2), a blue-light sensitive non-specific cation channel, activation of which results in depolarization...optogenetics. Successful stereotaxic surgery relies on identification of the bregma and lambda sutures and subsequent...bregma which allows you to quickly and reliably locate your injection site. We recently purchased this... fluorescent protein, which allows for easy identification of opsin expression, extent of viral spread...

...PMID: 18295576. PubMed Central PMCID: PMC4142810. 3. Mitalipov, S. and D. Wolf, Totipotency, pluripotency...potential of muscle stem cells. Dev Cell, 2001. 1(3): p. 333-42. PubMed PMID: 11702945. 6. Cowan, C.A....stem cells are: Their ability to self-renew or replicate while maintaining the undifferentiated “stem” ...Unipotency: As the name suggests, cells under this category can differentiate into only one cell type. Muscle...differentiate only into myogenic precursor cells [5]. Classification of stem cells Stem cells are classified into...somatic stem cells, these multipotent stem cells are located in small quantities in specific “stem cell niches...body. These generate specialized cells for their location, mostly to maintain cell numbers or replenish ...