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...Rab11, and Rab35 to understand their roles in forming the cilia that help establish left-right organization... and limitations – for example, microtubule-destabilizing drugs are impossible to target to specific subcellular...available as lentiviral plasmids if you want to make a stable cell line. This design allows highly localized,...reversible microtubule severing and has very low dark-state activity and toxicity. All of that means this light-powered...dissect the roles of three Rab proteins during a key stage of zebrafish development (Aljiboury et al., 2023...After 30 minutes of blue light illumination, substantial PA accumulates in the plasma membranes, indicated...moderately-enhanced 10x and 30x SuperPLDs avoid dark-state activity and cytotoxicity better than the 50x and...

...suspenseful. 4. Start preparing your talk early - like today Don’t wait until you need to speak to start creating...’s attention will start to wander when you are supposed to be done. They will start looking at their phones...future boss in the room, so it is a good idea to start thinking about public speaking early and often. ... audience is that it can help you avoid making mistakes. I once made a joke about social science to an...Describe how you pushed to a solution or overcame obstacles. Take time to consider the order of your narrative...t be hard on yourself if you stumble or make a mistake. Move on, the audience doesn’t really care about...

...how CRISPR can take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and...ART-induced viral control found in most HIV-1 patients. Obstacles for CRISPR HIV-1 therapy Both of these methods...survive, but preventing re-infection would require sustained expression of Cas9/gRNAs, which could result in...enough T-cells could be reached to ablate a well-established viral reservoir. A second challenge for CRISPR...binding. It’s also possible that HIV-1 may evolve resistance to CRISPR therapies through PAM or seed sequence...multiple drugs to lower the odds of developing resistance. Shortly after Kaminski et al. published their...not ablate viral function, but rather promote resistance to CRISPR/Cas9. While these results are discouraging...

...to put together DNA fragments. As a fun way to start, I highly recommend watching this entertaining video...vector of choice as shown in the figure below. To start, you need to have DNA fragments with regions of ...not work well if the ends of the fragments have stable single stranded DNA secondary structure, such as...Gibson assembly isn’t ideal for synthetic biology standards, which rely up on the re-use of parts between ...One way around this is to use a combination of standard sequences with overlap regions, such as in the...to cleave the 22kb vector. When followed by the standard Gibson assembly techniques described above, this... to get mixed up with Gibson assembly (they all start with a G!) but actually work by significantly different...

...antibiotic resistance cassette and you aren’t introducing an additional antibiotic resistance cassette ...where the inserted DNA includes an antibiotic resistance marker. After transformation, plate the cells...into the plasmid backbone contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing...cells carrying plasmids that contain a kanamycin resistance cassette will grow. However, often times, the...often used for cloning in yeast since antibiotic resistance spontaneously arises in yeast. For example, Markus...occurring genes for cloning purposes (antibiotic resistance genes, toxin genes, and the sacB gene). By tapping...

...that efficiencies of stable gene transfer using SB100x were comparable to stable transduction efficiencies...enzyme.2.) The transposase enzyme is expressed (red stars) and binds the inverted repeats (green); and an ...determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature...This new tool kit allows the rapid generation of stable transgenic cell lines which can robustly express...Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. Biotechnol J. 2015 Feb 4...hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun...

...collaboration for gene classification This inquiry started when Neil Thompson, a research scientist at MIT...at the time, got involved. Along with Philipp Pfingstag, a visiting graduate student from TU München at...was moved exactly as is from one organism to a distantly related organism, the gene may not express well...not rare codons and many genes where there are instances of rare codons, sometimes intentionally modulating...modulating expression in ways we don’t quite understand yet,” Kunjapur says. Percent sequence identity was...showed a growing trend of sourcing genes from distantly related organisms. In 2006, less than 1% of the...  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows ...

...Scintillon Institute, Allele Biotechnology, Florida State University, Indiana University, and the Karolinska...protein wavelengths, it can be imaged either with standard green fluorescent protein filters or yellow fluorescent...brightest teal fluorescent proteins that were photostable and had a high 480/530 nm emission ratio. They... in signal relative to mCerulean imaged with a standard CFP set. mTFP1 can be used as a FRET donor to ...CFP484. Collaborating with scientists from Florida State University, they used a mutagenesis approach and...mWasabi is 1.6-fold brighter than EGFP and is photostable. It also has narrow excitation and emission peaks...Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein...

...bacteria have just reached stationary phase, but the cells have not started to die off. Have any questions..., 2020. Now that we know all about antibiotic resistance genes, let’s consider another basic element of...cell. Figure 1: A plasmid map showing the standard features of a plasmid. So, how do you choose...researchers can ask themselves a few questions to get started: Will the plasmid be used exclusively in E. coli...will compete for the same machinery, creating an unstable and unpredictable environment. As a rule, plasmids... subculture directly from glycerol stocks, agar stabs, or liquid cultures. Freshly transformed cells also...

... and conditions to understand exactly what your antibody recognizes.  Understanding how the available ...the protein folded, changing it from its native state to a less folded, or denatured, one. This enables... protein of interest while it’s in a transition state or specific configuration. So how do you ensure ...protein as it exists in your assay?  First, try to understand, to the best of your ability, what factors may...during your sample prep process, imagining how it starts and if any steps could affect its configuration...assay that uses similar sample preparation - for instance, if you cannot find an antibody validated for ...

...don't, of trying to understand your completely unexpected findings, or understanding why your experiment...nothing if not solving your way through a series of obstacles in pursuit of achieving knowledge. Or, as we in...result of competing desires or (3) the presence of obstacles that need to be overcome. Conflict, and its inherent...them want to know more. This could be by outright stating the big-picture problem, by asking your audience...provide enough information so that the audience can understand the conflict. A science-based conflict gives ... and they all can be effective in the right circumstances.   However, in the midst of all these storytelling...

...alternative” anymore.   Since most scientists start out in academia, it is especially important for ...article well worth reading on this topic, the authors state, “Both explicit and implicit aspects of today’s ...”.   In fact, there are a lot of good reasons to stay and good reasons to go.  Here are some questions...calendar Academic researchers must be extreme self-starters and especially good at self-promoting—No one is... is going to tell you to pick up the pace Constant worry about grants and renewals What is different ...about something like this without sufficient data? Starting to learn about the diverse options and how to ...Science Careers How to Negotiate a Successful Lab Start Up Budget Read Our Management for Scientists Series...

...sales training and knew I had to start somewhere. I knew if I started with the Harvard Business Review... be to start reading a broad simple overview, learn some jargon and key phrases and understand enough ...and cloning, I decided to pack it in. I had just started month 8 of my 3 year Post-Doctoral contract at ...interview. The simplest place I could think of to start was “Marketing for Dummies”. The fastest way to ...academia, sales and marketing can be great places to start. Depending on what type of career you would like...

...system is to create stable transgenic cell lines. The traditional method of making stable cell lines is to...inserted into the genome fully intact, while, with a standard DNA plasmid transfection, a double-stranded break...with any integrated transgene). To overcome this obstacle, a positive selection cassette can be incorporated...containing a selection marker is included in a standard homology directed repair template to facilitate...use of negative selection largely overcomes this obstacle. Furthermore, we have found that transfecting ...Senior Scientist at Transposagen. He has a long-standing interest in molecular and cellular biology and.... Cooney, N. Malani, T. Brady, J. D. Sander, J. Staber, S. J. Wheelan, J. K. Joung, P. B. McCray, F. D...

...loci in the genome and the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two...CRISPR-based loci visualization tools by increasing gRNA stability. First, they began with the Broccoli aptamer system... loop motif within the gRNA) resulted in a more stable gRNA than when they inserted the aptamer at the...through subsequent design iterations to increase stability and minimize misfolding and recombination during... visualize three loci and measure inter-locus distances. They tested this system out on chromosome 19 ...IDR3) as a reference loci to measure inter-locus distances of loci pairs from intergenic regions, intronic... the National Academy of Sciences of the United States of America 112 10 (2015): 3002-7. PubMed PMID: ...

...in a single day -- all while maintaining social distance.  From front to back: David Gordon, Jeff Guo, ...coronavirus We are just at the beginning of understanding how SARS-CoV-2 interacts with human proteins...replication. While we and our collaborators are starting these experiments, there's so much to do that ... to fight the new Coronavirus. To help get you started, here are a few key details. Our set of constructs...Additionally, they have an IRES-Puro marker to facilitate stable cell clone selection. Expression is driven by a...been quite the administrative headache receiving constant email requests and tracking their fulfillment ...follow us on Twitter (QBI_UCSF and KroganLab), Instagram, and Facebook. Find more COVID-19 and coronavirus...

...will find a bike kitchen on the first floor and upstairs you will find our fully-equipped laboratory for... in the space and decided to reactivate it. We started out by holding a few journal club events and soon... BosLab inc. as a nonprofit organization in the state of Massachusetts. Because we’re conveniently located...equipment and safety training prior to getting started. For example, Timothy Stiles joined BosLab last... entrepreneurs using our space to perform early-stage experiments. For example, Dave Grawoig is using ...Community projects at BosLab Not all of our members start out with personal projects in mind. Thus, to encourage...Recently, Timothy Stiles and Mrina Nikrad have started the Open Yeast Engineering Project (OYEP). Through...

...be on your feet constantly in a lab? Will regular travel be required or will you stay in one location...Manager Patent Lawyer Science Market Research Statistician   Attention to detail Patience, accuracy ...including: Science Reporter Technical Writer Statistician Bioinformatician Science Writer Grant Writer...the number of available roles). Make your resume stand out Before you begin to apply for a job, it is useful...think of other aspects of yourself that make you stand out. For every role you need to show that you have... skill set and done your research, it’s time to start sending out applications. Check out our other career...

...neurons are physically connected, we can begin to understand how the brain works. It’s kind of like “the head...in a powerful way. Using brain structure to understand brain function So far scientists have identified...itself across these regions is important for understanding how the brain functions. The basis for these...neurons are physically connected we can begin to understand how the brain works. There are two methods of... spinal cord (Wang, 2018, Metcalfe, 2022) and substantia nigra (Tervo, 2016). Discrepancies in transduction...such as those in the spinal cord. Now that you understand this new tool, go forth and trace some neurons... Additional Resources on the Addgene Blog Get Started with AAV Learn more about Lentivirus Find out More...

...Piggybac transposon to stably integrate a reporter gene that contains a puromycin resistance gene followed by...prevent alternative start site initiation, the lab removed the native methionine start site. This first ...CRISPR reporters in the near future.  SRIRACCHA (stable, but reversible, integrated reporter for assaying...:  A donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual...Pruett-Miller SM, Blanc V, Davidson NO, Madison BB (2017) A stable but reversible integrated surrogate reporter for...