...tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community. Don E...PiggyBac transposons, the Woltjen Lab compared different variants of the polycistronic cassettes and discovered...is a new member of this class that works via a different mechanism - the PhoCl protein is irreversibly ...“caged” by fusion to specific domains thereby rendering them inactive in the cytoplasm. The authors use...interest. NHEJ- or HDR-induced mutations in ATP1A1 can render cells resistant to the small molecule ouabain -...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ...examining genomes possessing CRISPR systems with apparent self-targeting. Two of these Lmo proteins, AcrIIA2...

... Conversations with children playing in the background became a normal occurrence as Addgene and our university...light of the need for basic research during the current pandemic, universities are showing an unprecedented...of Illinois Chicago                     Boston Children's Hospital                           California...scientists. This partnership complemented each other's strengths: Ginkgo Bioworks’ infrastructure allowed them ...

...year bioengineering student, to Addgene. I am currently working on the QC/QA team as a DNA Sequencing ... food service industry, which followed a very different schedule. Fortunately, I was provided with a detailed...gave me enough time to become familiar with the different types of issues that were important to this function...Specifically as a co-op, it is encouraged to explore different teams and work on projects there. At this point...point, I have seen several lab processes and am currently working on projects for the Science Communication...

...neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists... populations using viral strategies Combining different approaches can increase the specificity of cell...optogenetic payload in neurons that project to different regions of the brain. A more locally defined set...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...share experimental design and data. With these different approaches to open science, we aim to accelerate...

...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three colors. The...specificity of these orthologs is key: due to differences in the PAM sequences required by each ortholog...specific for telomeric sequences and showed that different fluorescently labeled dCas9 isoforms are efficiently...target sequence. They were also able to label two different pairs of chromosomes using gRNAs specific to sequences...Addgene are excited to see what’s coming next! References Multicolor CRISPR labeling of chromosomal loci...

...trained to be good at learning new things—this is a strength you can draw on and demonstrate in your application...be excited to be there. If you are appying for diffrent types of positions (for example, bench research...resume should also be customized to highlight different skills and to get the important information in...hiring manager. Learn to confidently describe your strengths in your resume and don't be shy about asking for..., you will never get the help that can make a difference. Having your resume forwarded personally gives...

...and insecticidal proteins to the development of renewable biotechnology products and biosensors for herbicides...Development of a bacterial expression system for renewable biotechnology products Dr. Idah Sithole-Niang,...database for this important agricultural plant. Her current research focuses on endophytes, bacteria, and fungi...effect observed in bacteria. Learn more about the different CRISPR applications in plants.     Transgenic ...helped us coordinate and promote the awards. References 1. Martinez-Klimova, Elena et al. “Endophytes...

... Substrate: Some luciferase reporters, such as Renilla luciferase (RLuc) or NanoLuc, require injection...together genes from gas vesicle operons from two different bacteria, Bourdeau et al created two constructs... 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles is the amount ... invisible to ultrasound, which allows for differentiation of arg1 and arg2 ultrasound signals. This also...and arg2 plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan...

.... Northeastern compiles a list of hundreds of different jobs for students to apply to, and after a grueling...marketing and social media co-op. There are a lot of different worries that come along with entering the workforce...and co-founder, approached me and asked about my current projects and personal interests. We chatted about...participated in the discussions, I began to note trends in the data, and from that, suggest changes to ...of my co-op. I had to do research on loads of different best practices, create an email strategy, and ...

... tissue, in vivo. Two of their constructs are currently available as Cre-dependent, EF1ɑ-driven viral ...have high selectivity for K+ over Na+ and rapid photocurrent kinetics. They demonstrate the use of HcKCR1...two-photon excitation around 1,040 nm, has larger photocurrents, and is more selective for K+ than HcKCR2, making... be used as a potent optogenetic silencer. The current work also proposes promising avenues for future...anti-c-Myc [9E10] (isotype IgG2a) antibody is currently recommended for use in western blot, but we are...

...antibodies, and more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, ...Nonetheless, this tag can be purified with several different resin types and is a fairly small tag that is ...  Affinity tags vs. other tags There are many different types of tags, including epitope tags (FLAG, Myc..., D. W., & Wingfield, P. T. (Eds.). (2001). Current Protocols in Protein Science. John Wiley & Sons, Inc...

...leader-specific. How is leadership different from good management? Aren't all career skills leaderhsip skills...because it emphasizes the importance of these very different roles. At any stage in our careers we may be in...you are not fulfilling the requirements of the current task. Most of us start as doers, some become managers...boss) that you are ready to try operating in a different role?  I think the secret sauce can be summed ...

...Cong worked with Addgene Senior Scientist, Matt Ferenc, to organize the CRISPR forum's most popular questions...double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will... like the wildtype Cas9 does. There are a few references for the double nickase system, including one ... Sanger sequencing). Click here for a useful reference.  More FAQs, CRISPR protocols, and gRNA design... and optimized for specific plasmids Links to different software to help you identify your gRNA target...

...due to genetic drift and the use of genes from different microbial strains. It proved to be impractical... least 96% sequence identity when detecting a reference feature. For a coding sequence feature that may... could miss a common feature if the sequence differences exceed the threshold. This issue is addressed...often come in closely related versions that have different properties. To prevent misidentification, we augment...way for SnapGene users to apply features from a reference strain to newly sequenced strains through our ...

...obstacles that need to be overcome. Conflict, and its inherent tension, is interesting. It’s what drives a story...actually from a completely unexpected finding in a different experiment, this tool was developed because a ...couldn’t progress without it… These details often aren’t included in scientific communications, but they...scientists doing the work.  And while these pieces aren't always appropriate for your formal scientific communications...doing and sharing science in the first place?   References and Resources More resources on the Addgene blog...

...cells. The replicons of plasmids are generally different from the those used to replicate the host's chromosomal...include ColE1, pMB1 (which comes in a few slightly different but well known derivatives), pSC101, R6K, and ...considerations. *Requires pir gene for replication (reference). **F1 is a phage-derived ori that allows for...chloramphenicol – make sure your growth conditions aren't working against you!  The culture inoculum ...clarified? If so please, post in the comments.  References Chatterjee A, Sun SB, Furman JL, Xiao H, Schultz...

...AAV. You’ve never worked with AAV before, but you aren’t going to let that stop you. Where do you start?...capacity between the ITRs is ~4.4 kb. ITRs are inherently unstable due to their secondary structure, palindromic...species or location. Promoters can also drive different levels of expression. For example, CAG is a strong...Next let’s talk about some of the elements that aren’t required parts of an AAV transfer plasmid, but ...specific expression of your gene of interest. References Haery L, Deverman BE, Matho KS, Cetin A, Woodard...

... our values of transparency, reproducibility, and community-building. The transparency begins with sharing...whether any given antibody is available through different companies (Bandrowski et al., 2016). It’s also... the antibody sharing landscape as well.     References Andrews NP, Boeckman JX, Manning CF, Nguyen JT... Improving Reproducibility through Rigor and Transparency of Experimental Methods. Neuron 90(3):434-6....

...previously hidden epitope (right). Created withBioRender.com.   Biological Processes  If you’re feeling...outside of the cell. The same protein may have different epitopes available if it is being selected out...controls in validation, as you may need to test different configurations and conditions to understand exactly... what a given protein is doing or changing in different conditions. Just remember to keep good records...when you’re building your antibody library! References and Resources More resources on Addgene’s blog...

...antibodies for tagging purposes. Different antibodies bind different protein sequences called epitopes...infamous frankenstein, was constructed by grafting different parts together. By combining two optimized elements...living cells. To make these scaffolds specific to different targets, the scientists fused them to sequences...to build a more robust set of frankenbodies. References Ewert, S., Honegger, A., & Pluckthun, A. (2004...