...Research in Australia I then travelled abroad to work at the University of Queensland in Australia, this was...companies which are reliant on contracts with clients is that, when those contracts fall through or are cancelled...entitled to a Work-Travel VISA, which means I can gain employment for up to 6 months in Australia or another ...position in your field of interest before you dive straight into a full-time position. For my part, I really...with individuals who had very strong beliefs in traditional medicine, especially when I knew that there was... cheap and easy treatment for a disease that traditional medicine couldn’t fight effectively. I had to...(SOP’s) and every item I tested was constantly tracked with barcodes and spreadsheets. It was great to...

...video abstracts identified. The journal Cell had the largest average view count per video abstract at 2,947...antibiotic prescriptions for respiratory tract infections. The video abstracts provide a brief introduction to ... in many disciplines has been a long brewing frustration for me. I read a variety of psychology, business...science communication through video     Video abstracts about research, especially published research,...Scott Spicer at the University of Minnesota helps illustrate this: “In terms of total view count and videos... views) rounded out the top four average video abstract view counts.” For researchers looking to garner...garner more attention for their work, a video abstract may be a useful tool for turning video views into ...

...AAV5, AAV8) enable different patterns of transduction and transgene expression across different cell types..., Rumpel S. Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5...expressed from your cell line of choice after transduction with this virus. While various Cas9 mutants ...classic Cas9 nuclease, which introduces double stranded DNA breaks, can be expressed from lentiCas9-Blast...single stable Cas9-expressing cell line and then transduce this line with lentiviruses expressing various...experiments and over time (as compared to separate transductions with a single-vector system). Tips for using...is used to attain cell-specific expression in transgenic animals, where cre expression is restricted to...

...the opposite strand). ABEs make an A to G change (or a T to C change on the opposite strand). Both are ...place on the free strand of DNA and converts a C to U without producing a double strand break. UGI = uracil...repair (HDR), the pathway for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared...are base transition editors: converting one purine base to the other or one pyrimidine base to the other...other. Figure 1: Base transition editors catalyze purine-to-purine or pyrimidine-to-pyrimidine base... indicate the change introduced to the opposite strand following base editing and DNA repair. Black arrows...BioRender.com. How does base editing work? Base transition editing requires three elements. They are, broadly...

...but it is for a scaleable lab. When you need to track the location, quality, growth, and legal status ... that everything is grown successfully, we must track and manage all of these steps. Unfortunately, biology...they needed to, they’d have to individually keep track of any failures and do troubleshooting on their ... it’s monumentally difficult without software. Tracking lab procedures also gives you access to hard data...t have a system through which she could easily transfer her instructions for dealing with the samples ...repeated. We also now have a good interface to begin tracking analytics data for lab procedures. With this data...work more efficiently. Get on that barcode gravy train Buy a barcode label printer. Buy barcode scanners...

...emocionados! Después de recibir la traducción, Addgene contrató a una traductora científica profesional, L. Denise...receiving the translation, Addgene contracted with a professional scientific translator, L. Denise Roche...CRISPR 101 ya está disponible en español! Esta traducción fue regalada a Addgene por Daniel Mejías Sánchez...herramienta de edición ahora es parte oficial de nuestra colección de libros electrónicos. Descargar CRISPR...tiene una gran audiencia internacional, incluido trabajando con distribuidores de todo el mundo para ayudar...Muchas gracias a: Daniel Mejías Sánchez, PhD, traductor  L. Denise Roche, PhD, MD, editora, ldeniseroche...101 eBook is now available in Spanish!  This translation was gifted to Addgene by Daniel Mejías Sánchez...

...purification of tagged transcription factors in mammalian cells and transgenic mice.” PNAS 100.13 (2003...and decarboxylases. These enzymes catalyze the transfer of carboxyl groups to organic acids to produce...lipogenesis, amino acid metabolism and energy transduction. Biotinylation is the general process by which...is responsible for this highly specific post-translational protein modification. Most organisms have less...the reporter signal and easier detection. This strategy is referred to as the Avidin-Biotin Complex (ABC...surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated cell sorting...to attach proteins or polypeptides to a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith...

...remove one strand of the siRNA and are directed by complementarity to the remaining single stranded siRNA ...activity or their translation disrupted through physical blocking of the translation machinery. In the...WISH) for visualizing expression of genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to... by T7 promoters to allow for bidirectional transcription to create dsRNA. Students can produce DNA templates...the temporal and spatial localization of mRNA transcripts in tissue. This is useful for labeling specific...these constructs, developmental biologists have to extract RNA from their organism of interest, use the RNA... gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating Gene function by RNA...

...into… Transcription units (TUs), which are functional units composed of multiple parts transcribed together...a protein that regulated its own transcription, or the transcription of another gene? Now you have a genetic...As a plasmid repository, we are no strangers to cloning. If you’re reading this, then chances are you...Modular cloning, or MoClo, is a systematic cloning strategy designed by synthetic biologists to perform large...hierarchical levels: individual DNA parts, transcriptional units, and multigene constructs. Let’s start...start by examining individual parts and transcriptional units. Individual parts are defined DNA sequences...assembled into larger functional units which are transcribed together. Image courtesy of Robert Hurt. ...

...Tethering dCas9 to the histone acetyltransferase p300 or the DNA methyltransferase DNMT3A allows researchers...affect transcription independent of their enzymatic activity. They interact with transcriptional machinery...on DNA methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA....Mattei AL et al (2018). Genome-wide tracking of dCas9-methyltransferase footprints. Nat Commun 9, 597. https...specifically edit methylation sites.  Step aside, transcriptionally active dioxygenase  When I started to work...by distinct proteins and result in unclear transcriptional consequences (Sprujit 2013). Enzymatic domains...reality, editing your target site is not as straightforward as expected. TET1 can have many effects independent...

... modified cells are transferred into an animal and the behavior of the transferred cells is followed. ...generating a massive amount of descriptive transcriptomics data. Custom CRISPR screens designed to target...pathway with many receptors, signaling molecules, transcription factors, and ligands. The presenter then showed...are lucky and find that a druggable gene has a central role in your system, you could majorly accelerate...smaller validation screen using selected gRNAs extracted from the same or another library. These different...in animal model systems. Use Green Listed to extract gRNAs that target your list of selected genes. ... design. With Green Listed you can also easily extract gRNAs for the same gene from several libraries,...

...non-target strand. The helicase domain is then activated, and Cas3 begins unraveling the double-stranded DNA ...cleaving the ssDNA strands. In this way, Cas3 eats away at both the non-target and target strands by making cuts...every so often. Cutting both strands ensures that there isn’t a template strand left behind that the cell... regions on the non-target strand. Occasional cleaving of the target strand is not as well understood ...bypassed by leaving out Cas3 — a tactic some CRISPR transposase systems utilize with type I-F Cascades. ... section for details). The newly formed single-stranded DNA (ssDNA) is then pulled through Cas3. After...implies — after nicking, Cas3 reels in the non-target strand of DNA much like a fisherman would reel in a big...

...three double stranded breaks are repaired by two rounds of MMEJ. This double MMEJ strategy looks very similar...genome edits. But have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated...primary methods for repairing DNA after a double-stranded break. HDR copies the sequence from a repair template...fashion, with insertions and deletions common. In contrast, MMEJ uses regions with 5-25 bp of microhomology...are chewed back to reveal homology, allowing the strands to anneal. DNA synthesis then fills in the gaps...5-25 bp) regions of homology flanking a double-stranded break yield precise deletion of the sequence between...homology arms comes more time spent cloning. In contrast, the short homology required by MMEJ can easily...

... area that demonstrate that 85% of leaving postdocs pursue a career outside the traditional academic silo... areas quickly Jack/Jane of all trades More sources of training and resources within the company ...acknowledged perks of scientific and technical training is that these educational paths prepare you for...in US Biomedical workforce will NOT go into “traditional” academic positions. More recently, I have been...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...perspective on other types of science careers as their training progresses.  In From Academic Solos to Industrial...explicit and implicit aspects of today’s postdoctoral training can directly interfere with a seamless jump into...

...and they had been shown to to transduce the retina but only transiently. However new viral vectors capable.... PubMed PMID: 29868583. PubMed Central PMCID: PMC5960719. 8. Trapani I and Auricchio A., 2018. Seeing... years ago when completing her PhD and medical training in Ophthalmology. Gene therapy was in its infancy...adenovirus (AdV) and lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of...due to its considerable immunogenicity and its transient expression. LV results in long term expression... packaging capacity is well suited for larger transgenes. Unfortunately it has limited tropism in the ...and the one used in Luxturna™, can efficiently transduce various types of retinal neurons. Nowadays a toolbox...

...column or an ultrafiltration concentrator. As the name suggests, you may also concentrate your prep to...mammalian cells can perform human-like post-translational modifications that bacterial and most yeast ...protein you need to produce.  The most common yeast strain used for recombinant antibody production is Komagataella...be produced through a single or dual-plasmid transfection, depending on whether the heavy and light chain...they’ve been separated onto two. You’ll first transfect your cells with your plasmid(s), and then wait...to the desired concentration using these columns. If you are conjugating your antibody in a downstream...Whatever you choose, you’ll want an accurate concentration to give your downstream applications the best...

... protein translation. Its genome consists of “early” (E) and “late” genes, and the transition between ...makes sense that vectorization strategies include creating an AdV transfer plasmid with the E1 and E3 (Early...sequence (transgene) instead.    Although the late genes are usually present in the transfer plasmid for... for any virus, we will see that they are very central determinants of how AdVs behave, and in fact, of...Borkenhagen et al., 2019). AdVs are non-enveloped double-stranded DNA viruses, with a linear genome between 26–45... (and therefore between early and late gene transcription) is demarcated by viral genome replication (...cells that are infected by the vector, only the transgene will be expressed, since the early genes are absent...

...professional development strategy  Build your own Professional Development Strategy to increase your competencies...Approved Trainer by the Grant Professionals Association, and a Certified Social Media Strategist through...the highest likelihood of funding and develop a strategy and projections around that likelihood. We provide... and regulations impacting grant management, we track grant opportunity deadlines, and we ensure that ... materials for publishers. When the community orchestra needed a grant writer, Deb volunteered. Through...consulting firm, a Twitter Chat, and then our grant training and mentoring. How do I become a grant professional...expertise in grant writing, grant management, grant administration, and grant evaluation are available through...

...30644390. PubMed Central PMCID: PMC6333924. Howerton, Alexis R., Alison V. Roland, and Tracy L. Bale. "Dorsal..., inhibiting, or modulating. Viral vectors, in contrast, are considerably more versatile and specific ... circular corridor apparatus using a camera or tracking software. Locomotion is a highly complex process...project to the NAc, where enhanced glutamate transmission leads to locomotion sensitization. (for review... Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC projections to the NAc and decreased...risky). Taking advantage of these characteristic traits, rodents can be used to study anxiety levels in...injected Cre-expressing AAV-5 into the DRN of a transgenic mouse harbouring a Cre-dependent IL-1 beta overexpression...

...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills...employee who's coming directly from grad school. The transition has been a bit rough for her (3 weeks in and ...there are any resources that can help with the transition.” The lab-to-desk-job culture shock is a real...scientist colleagues have as well. Making the transition from the bench can be jarring for the unprepared...resources out there for scientists making this transition. In this installment of the Addgene career guide...formulate hypotheses and conduct experiments. You transfer liquids between tubes, look through microscopes...What should you expect during the lab → office transition? I’ve already provided a few counterpoints to...