...Diagram of AAV components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that...primary application of AAV is delivery of genetic cargo to a host cell. In order to make room for a gene... (rAAVs) with maximum “trunk” space for genetic cargo. Typically, when researchers speak about AAVs, they...switches placed in your cell type of interest. Careful combination of promoters, gene switches, and/or...second strand of DNA before expression of their cargo can begin, which all takes time. Though the time...standard ballpark estimation. Once expressed, viral cargo is usually quite stable and in some mammalian tissues...self-complementary AAV (scAAV) may be an option (McCarty et al., 2001). This AAV is designed so that the...

... Laboratory. Are you a science student or early career researcher looking to break into science communication...Everybody goes about this in their own way. The career paths into science communication are as varied ...strategies will get you started:  Subscribe to Career Blog Posts from Addgene 1) Twitter – Get your voice...and science blogs like @WIRED Science. Don’t be scared to engage with them. Comment on their activities... like Ed Yong jumpstarted their science writing career by blogging. Writing a blog is not only a great...know and get known by editors and others whose careers are in advance of yours, but also get inspired ...Imperial College London. Similarly, the University of Cardiff offers an MSc in Science, Media and Communication...

...characterized: mScarlet (fluorescent lifetime of 3.9 ns, quantum yield of 0.70), mScarlet-I with T74I (...compared to mScarlet, fluorescent lifetime of 3.1 ns, quantum yield of 0.54) and mScarlet-H with M164H...compared to mScarlet, fluorescent lifetime of 1.3 ns, quantum yield of 0.20). All three mScarlet variants...New bright monomeric red fluorescent protein - mScarlet! The Dorus Gadella lab has recently developed ...synthetic red fluorescent protein variant they call mScarlet. Bindels et al used a combination of deliberate...protein fusions. The Gadella lab has deposited all mScarlet constructs with Addgene. Bindels DS, et al. Nat...Methods. 2017. PubMed PMID: 27869816 Listen to our mScarlet Podcast Segment    Photocleavable protein PhoCl...

... are ways to create scarless mutations, ones that don’t leave any undesired scars in the chromosome. These... Chris R., and Kristala LJ Prather. "The no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome...an antibiotic resistance cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression...CRISPR-Cas9 technology has also aided in the creation of scarless mutations, both in stand-alone systems (Jiang ...replication to allow replication only in hosts carrying the bacteriophage lambda pir protein. This allows... Allelic exchange is a great technique for the scarless modification of bacterial genomes. It can accomplish... New Suite of Allelic Exchange Vectors for the Scarless Modification of Proteobacterial Genomes." Applied...

...serve you well. The happy byproduct might just be a career opportunity. The focus of this article will be...from. Some of the best talks I have seen by early career scientists are those in the Young Scientist Seminar...this opportunity to recommend all the science and career talks at iBiology which hosts an amazing amount...amount of useful, entertaining science lab, career, and policy content. Watch Joanne give her "Not" Networking...will be listening.  Are they early, mid, or late career?  Will many non-scientists be present?  Always ...when the host told me and wished I had been more careful. 3. Tell a story We are biologically programmed...about something you know a lot about and that you care about. Don’t try to present topics that are unfamiliar...

...Will I leave academia?” and "What type of science careers would would make a good fit for me?" Whatever would... headlines entitled “Presentation Experience”, “Career Achievements”, and “Academic Achievements”. Leaning...had published with and whether I would be able to carry out a yeast-2-hybrid library screen; instead I thought...Has this person been consistent throughout their career?”, "Have they sought out opportunities to expand...marketing knowledge. For most PhDs looking at careers outside of academia, sales and marketing can be...great places to start. Depending on what type of career you would like to build, sales and marketing gives... more posts about Scientific Career Development ...

...behind. Furthermore, piggyBac™ offers a large cargo-carrying capacity (over 200 kb has been demonstrated...direct integration of the donor-transposon (carrying your cargo of interest) into random ‘TTAA’ sites throughout...gene modification system and the scarless, Footprint-Free™ removal of cargo with the excision-only piggyBac...throughout the genome. When using piggyBac™, the cargo is inserted into the genome fully intact, while,...transfection, a double-stranded break may occur in your cargo. Additionally, with piggyBac™, the number of integration...utilizing a piggyBac™ transposon in a donor plasmid carrying the DNA edit of interest, along with a site-specific...

...Addgene Read Additional Career Posts on the Addgene Blog Browse Our Careers Page to See What Positions...20’s, new graduates often ask me for advice on careers available to new science graduates and the pro’...enjoyed some aspects of the work, such as physically carrying out experiments like PCR and running gels, but...and other reasons, I started  looking for other career options after 6 months. Returning to university...hands-on lab work for a dissertation. I chose to carry out my dissertation research in a lab in California...virology testing company. This particular company carries out clinical trials of drugs and treatments for...also understand my job would not suit everyone. My career path may seem quite unusual but it has allowed ...

...permanently integrate a genetic cargo into a cell. These viruses have a cargo capacity of ~ 8 kb, ample room... transient expression of a genetic cargo or have a very large cargo. Fast fact: All viruses discussed...itself to direct application of virus. However, the cargo capacity of AAV is ~4.7 kb, almost half that of ...doesn’t require genomic integration to express its cargo, in fact its viral life cycle does not include genomic...integration. While the expression of adenovirus cargo is high, it’s generally not as long lasting as AAV...some adenoviruses pack up to ~35 kb of space for cargo! Adenovirus is an effective option if you need high...

...business card – even students and postdocs. There are many inexpensive/free places to order cards. There...good reasons to have a card but two of the best are: if you give someone a card they are more likely to...meeting other scientists is not a priority for your career. Actually, it is crucial for all scientists, academic...you if you follow up and if you give someone your card, they have to give you theirs! Think about how you...traveling with a poster roll, say hello. Keep your cards handy and make sure your name tag is always visible...webinar with Joanne Kamens via ASCB Check out more Career Advice blog posts to continue improving your networking...

...protein (mScarlet) with either a seven or eight amino acid linker between them. The Ace-mScarlet voltage...results in higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive...inhibitory rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET. Image ...created two plasmids containing a red-shifted Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin...blue-light-sensitive channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et...

...Identifying Career Pathways for PhDs in Science”, through which she collects data on the careers of science...Resources at Addgene Learn More about Careers in Science in Our Career Posts! Get Advice on Networking at...academia may in fact be just another ‘alternative’ career.   The news is not all bad, however - there are... in STEM fields (6). Melanie Sinche is a STEM career counselor, Director of Education for the Jackson...that biomedical graduates take in their future careers. To participate in the Future or Research Symposium...Sauermann, Henry, and Michael Roach. "Science PhD career preferences: levels, changes, and advisor encouragement...

...across the membrane of cardiac cells. Similarly, neuronal signaling is carried out by action potentials...: Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin Ace2N and mScarlet. Ace2N... package your genetic cargo in lieu of their own genomes and deposit this cargo upon host infection. Packaging...directly related to the electrical impulse of the cardiac cells. To study this phenomenon you will likely...Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from Beck et al, 2019 under CC BY 4.0...

...my scientific expertise. Organizing a career fair for early-career scientists provided a crash course in...of the the skills favored by research-intensive careers. External collaborations are becoming increasingly...a message asking how things are going. Science careers where the ability to start and maintain external...is important for success in research-intensive careers. In addition, this skill is a hallmark for jobs...Additional Resources on the Addgene Blog Check out Our Career Topic Page Read Other Posts in the Transferable...Addgene.org Check Careers at Addgene ... science career...

...new FPs - the mCarmine story Article contributed by Caroline LaManna  Listen to the mCarmine podcast segment...by Klaus Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains...tested a series of mutations. Through this process mCarmine was developed (mNeptune684 with C65S, T107K, A108V... 603 nm, while the emission maximum is 675 nm. mCarmine is monomeric, 4-5 fold brighter than mNeptune684... test out this new FP in your own experiments, mCarmine is available in a mammalian expression vector ...

...daughters playing with some cards and I had a quick flashback to time spent playing a card game called “Magic”...this game was played. These rules restricted which cards could be played when and, depending upon who your... 100% student engagement. I figured, an engaging card game could help me get the engagement I wanted. ...disease case. Then, each team received a whole set of cards (Promoters, Inserts, Tags, Origins of Replication...each element to use, they had to place the selected card on the board game. They had 35 minutes to develop...and some were so enthusiastic that they took the cards and the board with them. My impression was that ...

...Assembly. Gibson Assembly allows the production of scarless DNA constructs using homologous regions to guide...the joining reaction. BioBrick Assembly will leave scar regions at the site of fragment joining, but this...use of Type IIS restriction enzymes results in scarless assembly. Finally, the NEBuilder® HiFi DNA Assembly...fragment overlap sections, while still creating scarless plasmid inserts.  Want to Learn More about DNA...reset and ligate mismatched overhangs, yielding scarless DNA assembly, and allowing fragments to be taken...used directly in DNA assembly will still yield scarless assemblies. Overview of NEBuilder HiFi DNA assembly...and primer design results in covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR...

...components to convert the cargo from RNA to DNA and permanently insert the cargo into the host genome. Lentiviral-specific...purposes. The primary plasmid with the delivered cargo (often a gene) is called the transfer plasmid while... most viral experiments is to deliver a genetic cargo, typically an expression construct, to cells/organisms..., though. Viral packaging and entry The genetic cargo requires viral packaging to infiltrate and integrate...functions to infiltrate the host cell and deposit viral cargo. Gene expression While AAV often does not integrate...inverted terminal repeats (ITRs) is the promoter and cargo, in that order. This will all be ssDNA within the...

...right path for me. I began exploring alternative career trajectories within the industry, but the transition...fulfilment in supporting my team’s development and career growth. This realization led me to pursue a shift...I discovered strategies to effectively pivot my career development and align it with these new goals, ...final choice Management is not a one-way path. As careers evolve, many professionals move between technical...that managing isn’t for you at this point in your career—and to step back into a technical role. My advice...transitions as an integral part of your broader career development.   Matteo Tardelli authored the book...metabolism. He now guides fellow PhDs in their careers with actionable steps and execution plans.   Resources...

...on Addgene.org Learn more about Careers at Addgene Watch out career videos  ...addressed in numerous essays and workshops (see the Careers essays in the journals Nature and Science, for ...what will be needed to achieve your intended next career stage and create a timeline for achieving these...designing and interpreting experiments, as well as career guidance. In my own experience, great mentors often...needed, this is a strong indicator that this person cares about your science and your development. Taken together...'ll be better prepared for your future intended career and you'll become a better scientist in the process...getting a faculty position Academic v. Non-Academic Career Decisions Check out our transferable skills guide...

...the available sgRNA-cargo constructs utilize a U6 promoter and contain the RNA cargo cassette inserted ...to observe intra- and post-Golgi segregation of cargo during their transport. Finally, the system can ...MluI-XhoI-SalI) chaining results in a translatable ligation scar (Val-Glu) between each module to permit easy construction...compared to other viral systems due to its large cargo capacity, minimal cytotoxicity, and broad host-tropism...and their collaborators in Robert Campbell’s lab carried out five rounds of random mutagenesis on a library...CRISPR-Display: A novel CRISPR technology for targeting RNA cargos to specific DNA loci John Rinn’s lab is interested...CRISPR-Display uses the dCas9 enzyme to deploy large RNA cargos to specific DNA loci. In CRISPR-Display, your RNA...

...Bacterial Colonies can be found below: Career videos Ever needed career advice or wonder what it’s like to...ll hear all about their career paths, future goals, and get some useful career advice.  Our first video...you to find content relevant to your research and career goals, or to answer any questions you might have...to work at a biotech nonprofit? In our Career Video series, we feature interviews with real Addgenies ...

...transposition, genetic cargos must be flanked by the L and R ends - thus, scarless insertions are not currently...efficient and programmable insertions of large DNA cargos into bacterial genomes. This new capability was...DNA (aka mini-transposon), which contains the DNA cargo of interest flanked by ~50-150 bp transposon end... which offer simple transformations for smaller cargos, and the ability to easily cure cells of the plasmid...spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid(s) into...programmable insertions of even very large genetic cargo, at one or multiple desired genomic loci. Many applications...

...found on the UK Career Site for Researchers called Vitae. Vitae recently published a career development framework...learned a lot. Whether you are pursuing an academic career path or moving in a nonacademic direction, there... put you in position to pursue a diverse set of career paths. Check out Joanne's Reddit AMA Here are some...defining characteristics of nonacademic science careers is that a team is almost always involved. Demonstrating... framework for career skills. Download a summary PDF here or try out the online Researcher Development...

...using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol 14:447–455 . https://...small RNAs in their respective species. Addgene carries >30 “empty gRNA” backbones which contain a plant...ubiquitin promoters (Belhaj et al., 2013). Addgene carries Cas9-containing plasmids for knockout, activation...CRISPR-Cas9 components into plants. Nanoparticles (carbon nanotubes, Kwak et al., 2019), DNA nanostructures...addition to the plasmids described above, Addgene carries several useful CRISPR toolkits for creating plant...Targeted delivery of nanomaterials with chemical cargoes in plants enabled by a biorecognition motif. Nat...

...including biotin carboxylase and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic...important cellular metabolites. Biotin acts as a carboxyl carrier from one compound to the other. These biotin...lysine in the entire organism on the biotin carboxyl carrier protein (BCCP). Discovery of the biotin-avidin...

...thinking about my career options is being an overthinker. Being an overthinker can make me scared of taking risks... no idea how to translate them into real world careers. I figured this internship would be a perfect way...was certain I would not end up in a life sciences career but life has a way of making unexpected decisions...future. They made me rethink having a structured career because our mindsets and needs can change. Having...say with certainty that Addgene is a company that cares - not only in terms of its employees but in general...

...pAAV-Ef1a-DIO mScarlet AAV5 Controls Karl Deisseroth New serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV5 Optogenetics... a whole host of (awesome) research not easily carried out before. We have a bunch of other new AAV vectors...AAV-GfaABC1D-Cre-4x6T AAV5 Recombinases Stanley Thomas Carmichael New viral prep pAAV-EF1a-fDIO-Cre AAV5 Recombinases...Ofer Yizhar New viral prep pAAV_hSyn-DIO-PdCO-mScarlet-WPRE AAV1, AAV5 Optogenetics Ofer Yizhar New...Edward Boyden New serotype pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE AAV1 Optogenetics Karl Deisseroth New serotype...

...including a Christmas Chemistry symposium and a career night. The society also visits high schools to ...become involved with. The society provides talks and career support for a range of different scientific disciplines...talks for both students and the general public, career events, and networking meetups. The society hopes...interested in this field excel and develop their career skills for positions both inside and outside academia...Resources at Addgene Read Our Posts on Networking Get Career Advice from Addgene Bloggers Learn How to Get Involved... career...

...Resources at Addgene Read Other Careers Posts Visit the Addgene Careers Page Learn about Management for...academia? Did I run screaming from the lab like the cartoon on the right?!? I would love to tell such a dramatic... a PhD and even more glad that I thought about careers outside of academia during my graduate training...you start thinking about preparing yourself for a career while still in graduate school, you’ll be well ... career development...