...demonstrate this quality.   I am often asked to give career seminars on Leadership Skills. After attempting...leadership different from good management? Aren't all career skills leaderhsip skills when done well? Do you...know it when you see it? “The Three Stages of Your Career: Doer, Manager, Leader” by Charlotte Beers, divides...these very different roles. At any stage in our careers we may be in a cycle of doing, managing or leading...spending a lot of time in doer mode even late in our careers.  Managers make sure the work gets done Managers...happy with a primary role of doer for your entire career. Scientists train as doers and often maintain an...

...two plasmids: pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries a spacer targeting the... of interest and kanamycin resistance E. coli carrying the phage recombineering machinery are first electroporated...only 2-3 hours incubation. E. coli and T. citrea scarless editing plasmids Sheng Yang’s lab describes a ...recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang et al., 2015...Gram-positive bacteria Jason Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting...10.1021/sb500351f Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG (2018) Synthetic CRISPR-Cas gene...https://doi.org/10.1128/mbio.00019-20 Singh AK, Carette X, Potluri L-P, Sharp JD, Xu R, Prisic S, Husson...

... Courses HHMI Resources for Early Career Scientists Science Careers: Running Your Lab (pdf) Advice for... next professional adventure begins. While many career pathways exist for scientists and engineers, a ...to take things slowly! It is more important to carefully select first hires who are well-suited, productive...for starting your academic lab, download Science Careers' "Running Your Lab" booklet. What has been the...Institute (HHMI) offers some great resources for Early Career Scientists, including their "Making the Right Moves...

...find those that do not carry a given dominant disease-causing gene, or that carry only one copy of a recessive...mutation in MYBPC3 that leads to hypertrophic cardiomyopathy. An adult male with a heterozygous 4 bp deletion... wild type allele (Figure 1). 72.4% of embryos carried two wild-type copies of MYBPC3 (versus 47.4% of...of untreated controls.) 27.6% of embryos carried indels at the mutation site, indicating that repair proceeded...many situations. In the case of hypertrophic cardiomyopathy and other genetic disorders, pre-implantation...

...Resources on Addgene.org Check out our careers page Watch our career videos ... school and beginning to research non-academic careers, I always felt that descriptions of transferable... this skillset. After sifting through dozens of career advice articles, I settled on defining problem ...to research and the lab, you may not be able to carry out the entire problem solving process on your own... and effectively will help you grow in whatever career path you choose.    Resources Additional resources...

...workshop, it is often “all hands on deck” to take care of the organizational logistics. Setting yourself...path to science policy and science communication A career in science communication or policy requires broad...currently in academia who are interested in such careers should take advantage of their academic environment... own blog. Even if you do not end up pursuing a career in science communication or policy, being able ...resources on the Addgene blog Check out our series on careers in science communication Find all of our science...Extracurricular activities for a strong science career path Resources on Addgene.org Learn how Addgene...

...have been derived from muscarinic receptors. The most popular agonist for muscarinic receptor derived DREADDs...cn500325v Goutaudier, R., Coizet, V., Carcenac, C., & Carnicella, S. (2019). DREADDs: The Power of the...Perlapine, and Olanzapine, that can also activate muscarinic-derived DREADDs. A ligand co-developed by Bryan....2016 Weston M, Kaserer T, Wu A, Mouravlev A, Carpenter JC, Snowball A, Knauss S, von Schimmelmann M, ...

...https://doi.org/10.1016/j.isci.2020.101330 Split-wrmScarlet, a fluorescent proteins system for Caenorhabditis... elegans? By using a split FP approach. Split-wrmScarlet is a new split red FP that is three times brighter...Calico Life Sciences. Like many split FPs, split-wrmScarlet is composed of two FP fragments that are not ... engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. ...with a ssODN template to introduce the small wrmScarlet11 which is less than 60 nt in length to tag the...distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different subcellular ...

...and pCutamp.  pDonor carries a customizable cargo gene in the form of RE-cargo-LE (RE, right end; LE,...transfer on transposition-inducing agar lead to higher cargo copy numbers. (D) The effect of gene dosage on N-acetylglucosamine...site-specific multicopy chromosomal integration of cargo without using screening markers. Capability to construct.... Ready for metabolic engineering with improved cargo dedicated to gene (cluster) interruption.     ...

...assemble up to 6 fragments in one step, resulting in scar-free assembly that does not require the presence...cloning, you would have to do it in two steps, and a scar would likely remain between the two fragments. However...tag sequences into the vector in one step without scars as depicted below. First, you need to design primers...assembling multiple fragments in one step with no scar sequences remaining in the final product. The long... no requirements for restriction enzymes, and no scar sequences between fragments. Some of these products...vectors containing these integration site and leaves scars between fragments, but allows for easy movement ...

... also been found to transduce skeletal muscle, cardiac, and retinal cells (Issa et al., 2023). AAV1 capsids...it has been shown to transduce skeletal muscle, cardiac, pancreas, smooth muscle, CNS, kidney, and retinal...efficiency, and is an excellent choice for targeting cardiac cells (Bish et al., 2008). It can also cross the...find experimental reports on commonly used vector cargo designs and administration doses. Don't forget to...Adeno-Associated Virus (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1, AAV6, AAV7, and...Wosen, J. E., Nagamine, C. M., Chapman, M. S., & Carette, J. E. (2016). An essential receptor for adeno-...

...non-homologous end joining (NHEJ), this will leave behind a “scar”, with nucleotides missing or added at the broken...CRISPR and find that many, if not all, the mice carry a frameshift mutation. For knock in with a donor...mice and are usually successful in deriving mice carrying the intended mutation. With a donor plasmid, the... In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA locus as assessed ... Jaenisch R (2013) One-Step Generation of Mice Carrying Reporter and Conditional Alleles by CRISPR/Cas-Mediated... Jaenisch R (2013) One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated...

...Advisor, Boss – who will help me advance my science career? We spend many years becoming scientists. It takes...or more for just the “training” portion of our careers. In that time we may have only 2-3 formal supervisors...guidance and experience. Navigating a fulfilling career in science can be challenging – is advice and guidance...researchers who joined a recent Twitter chat for Early Career Researchers.  Many of the participants had certainly...such a dedicated person guiding the start of their career. “I want to be a PI that holds my students accountable...

... depositors who are at various stages in their careers to get advice on choosing a lab, picking your research... anti-tech or pragmatic.” Advice for pursuing careers outside of academia Recently, graduate students...students and postdocs have become more aware of careers outside of academia. How do PIs assist their mentees... happy, and match their personal goals to their career choices.” She points out that a PhD teaches analytical...thinking skills that are applicable to a variety of careers outside of academia. She has had previous graduate...

...transposase. It has a cargo capacity >100 kB, although integration efficiency decreases with cargo size. Sleeping...cells without decreased efficiency, with a maximum cargo capacity of ~200 kB of DNA. Similar to piggyBac,...for AAV and ~8 kB payload for lentivirus). Large cargos, such as the ~11 kB cDNA for dystrophin, the gene...donor DNA or mini-transposon that contains the DNA cargo of interest flanked by ~50-150 bp transposon end...to allow for the targeted delivery of large DNA cargos (>100 kB) to specific locations in the genome. ...

...or seemingly unrelated ideas. It's beneficial to carry a notebook at all times (or use the notes app on...invaluable for me, allowing me to pinpoint areas of my career and life where I excel (and thus feel grateful)...adaptability, a crucial skill for a successful research career, whether within academia or industry. Albert Einstein...metabolism. He now guides fellow PhDs in their careers with actionable steps and execution plans.    ...resources on the Addgene blog Experimenting with New Careers While in Grad School How to Lead a Great Meeting...

...North Carolina Research Triangle area, scientists give talks and science cafés at the North Carolina Museum... others working in your own field. Alternative careers are now the norm for those (like myself) with a...his friends and family finally understood why he cared to study mice on some distant island. He got a little...  If you are serious about science writing as a career, the Science Communication Program at the University...

...started to become very aware of how little care I took with my discards and to be mindful of the lab materials... use plastic bags.  We also try to recycle our discards according to the Cambridge, Massachusetts guidelines... to be used for 3 months before they could be discarded.  Seeding Labs does an amazing job shuttling unwanted... downsides—there was no cover so you had to be careful not to stick your fingers in during a run or risk...

... genetic cargo — the viral genome — is swapped, at least partially, for a nucleic acid cargo of your choice...virus! The constructed viral vector containing the cargo of interest is then typically transfected along ...genome has been swapped for alternative genetic cargo, they can’t reproduce more functional viruses later...achieve? For example, do you want the nucleic acid cargo to be integrated into the host genome? Expressed...

...struck with the realisation that I didn’t want a career in neuroscience—whatever that entails. Despite ...decision I had unwittingly begun my journey towards a career in science communication, an established field ...required of me for work, I would have a high level of career satisfaction indeed.   For many venturing forth...research. In my case, an interest in neuroscience was carried over and allowed to percolate within a new intellectual...thesis in science communication, I had the chance to carry out a comprehensive review of the literature informing...

...exchanging business cards for awhile or ever so your LinkedIn profile link can be your new card. Follow your ...about building genuine relationships with people you care about knowing. How do you follow up with new connections...someone who you need to speak to for informational or career purposes.  Don’t only talk about work or ask for...to a job in pharma and she followed an academic career path. Through a twist of fate, we got back in touch...

... 20] Osteoclasts and osteoblasts [21, 22] Cardiomyocytes [23, 24] Smooth muscle cells [25] Skeletal... recent works, transdifferentiation is usually carried out by expressing transcription factors specific...vitro methods are similar except that the vectors carrying the transdifferentiation factors are directly ...reviews journal. She joined Cell Press to pursue a career in science communication and publishing after completing...al., Chemically defined generation of human cardiomyocytes. Nat Methods, 2014. 11(8): p. 855-60. PubMed...PMCID: PMC4169698. 24. Lian, X., et al., Robust cardiomyocyte differentiation from human pluripotent stem ...

... Novartis researcher, Carrie Bhang. The ClonTracer Library, deposited by Carrie Bhang, a research investigator...sequencing (NGS). The library was developed when Carrie was a post-doc in Frank Stegmeier’s lab in Novartis...put the ClonTracer library to use! Many thanks to Carrie and Novartis for taking the time to talk with us...this powerful resource to the Addgene community! Carrie Bhang is a research investigator in In Vivo Pharmacology...

...idea to move around to different labs during your career; you learn new techniques and interact with new...perhaps in a different country, choose where you go carefully. Funding agencies want value for their money so...fellowships. Also, if you were awarded money to carry out research, be sure to make it clear what the ...occasions. Strategize, choose calls that suit your career stage / research area, and focus on producing a... Become a Grant Writing Professional Facilitate Career Planning and Personal Development with an IDP Get...

...and animals. Nanoblades can also carry different types of CRISPR cargo including RNPs for indel formation...gRNA and Cas9. An “all-in-one” Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can ...target cell membrane, and transfer their CRISPR cargo into the zygote. As a proof-of-principle, the team...stealthily delivery many types of CRISPR editing cargo to difficult to target cells. When compared to existing...

...a retroviral system this looks like the genetic cargo in one plasmid (the transfer plasmid), the envelope...onto distinct plasmids for safety.    Viral cargo-specific safety   After replication competency, ...construct is being delivered, that is a higher risk cargo for a researcher to come into contact with than ...than GFP. Similarly, if the viral vector carries an shRNA to a gene essential for cellular function, this...integrating viruses – they physically deposit their cargo into the host’s DNA. These viruses also have a propensity...

...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...your virus and assaying for expression of a gene carried on the transfer plasmid or quantifying the number... low levels of virus and can be used to track carryover. Functional titer Functional titer is considered...This method is particularly useful when vectors carrying fluorescent markers are used as it eliminates ...approach utilizes the antibiotic resistance genes carried on transfer plasmids. Target cells are transduced...

...negotiation skills that would be useful in many careers. I learnt that the overall voice may not always...postdocs and graduate students to think about their career options, gaining transferable skills such as leadership...assist you and make you more qualified for future career opportunities. Becoming involved is not too difficult...JulietAdvocSci.        Further Reading Find more Career articles on the Addgene Blog Read Joanne Kamen'... career...

... delivery to the brain. The AAV (a) capsid, (b) cargo, and (c) delivery method determine cell type- or...or retrograde transport in neurons. (b) Genetic cargo can be customized to control transgene expression...have many of the same restrictions, like limited cargo space (~4.7 kb) for your genetic payload — there...miRNAs, enhancers, and switches all take up precious cargo space in AAV vectors. Dual expression systems, which...published systemic AAVs to find commonly used vector cargo designs and administration doses. Note: If capsid...AAV technology for delivering genetically encoded cargo to the nonhuman primate nervous system. Current ...throughout the central nervous system. Nature Cardiovascular Research, 1(4), Article 4. https://doi.org/...

...Like any major life decision, the transition felt scary. While I knew two people from graduate school who...March, and the jobs are mixed into the general careers section of their web site. Each position is for...opportunities to help you with the next step in your career.   Many thanks to our guest blogger, Laura Desrochers...Additional resources on the Addgene blog Check out all careers related blog posts here Find 9 tips for a successful...Addgene.org Find open positions at Addgene Check out our career videos ...