...Creativity is required to develop new projects and test novel approaches, a necessary skill set for any ...be easy to optimize virus production with roller skates—though I could get around the lab faster that way..., easy goal. For example, set a timer for five minutes and come up with as many solutions as you can in... is simply to throw out ideas.  When your five minutes are up, if you haven’t come up with any ideas obviously... or even very frequent, but establishing a few minutes every month dedicated to thinking about the future...time crunch, your break can be as simple as a few minutes of meditation or a walk to the break room for a...

...simultaneously. Image from Li et al., 2021.   The updates involved the following changes. In pCas9, we 1)...the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA. The ccdB serves...32 hours instead of ~60 hours. We successfully tested the pEcCas/pEcgRNA system in 4 E. coli K-12 strains...strain, and 1 Tatumella citrea, with a total of 70 sites edited. We optimized the strategy for plasmid curing... into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the plasmids by targeting...Chinese Academy of Sciences. He joined Shanghai Institutes for Biological Sciences in 2000 and became a ...

... suitability for in vivo studies. Testing AAV-SaCas9 in vivo To test the efficiency of AAV-SaCas9 in vivo...Cas9 genome editing varies across targets, they tested two genes in mice. For both genes, they saw indel...polypeptides that, when at the ends of two proteins, mediates protein trans-splicing, similar to an intron in...proof-of-concept split intein SpCas9 which had modest editing rates in HEK-293T cells when compared to the full-length...genes in human cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038/srep10777...

...activated” Cas13 produces a quantifiable signal that indicates the presence of your targeted nucleic acid. To...commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the presence of your ...for using SHERLOCK to detect SARS-CoV-2 RNA. The test is started using RNA purified from patient samples...Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends after dsDNA breaks. The...per second. A quantifiable fluorescent signal designates the presence of your DNA of interest, in this...CRISPR-based platform on which an infinite number of tests” for biosensing can be built upon. Mammoth Biosciences...

...protein. Such flexibility makes aptamers ideal candidates for biosensors. Aptamer biosensors are used in...oligonucleotide-only sensor could save precious minutes in a time sensitive detection assay. In 1999, ... selection of Spinach with a target metabolite creates an aptamer that will only bind to DFHBI in the ...dependence. Though initially still unstable in eukaryotes, the Jaffrey lab has since developed new RNA-...RNA-based scaffolds to contain and stabilize Broccoli cassettes, thereby extending their use into mammalian cells.... The nature of aptamers makes them excellent candidates for developing assays for otherwise difficult-to-study...

...sterile due to a failure of primary spermatocytes to enter meiosis. Testis of cookie monster mutant flies ...component of the fly actomyosin cytoskeleton that regulates cellular contraction. This network is composed...the formation of the visceral mesoderm which differentiates into the muscles of the gut. Embryos that lack...bagpipe: two homeo box genes that determine cell fates in the dorsal mesoderm of Drosophila." Genes & Development... "Myosin light chain–activating phosphorylation sites are required for oogenesis in Drosophila." The Journal...

...chalk talk." However, few, if any, faculty job candidates have seen an actual chalk talk. Their first exposure...session lasts up to an hour. You will get 5-10 minutes to present your research vision uninterrupted and...begin working through your aims over the next 45 minutes. The faculty will ask you lots of questions. Your...Comparing model A vs. B is more constructive than testing the validity of a single model. The first approach...is: What is the big question or mystery that motivates your research? This is the central idea. It should... Jennifer Lippincott-Schwartz at the National Institutes of Health, was an Associate Professor at Albert...

...you realize events are plotted on an invisible Cartesian plane (right.) Each event is plotted by the amount...different population.  Gating and plot orders  As the gates progress from broadest markers to most specific,...conjugated to fluorescent markers are used to identify monocytes (upper right) and a mixed population of B cells...understand how the researchers decided to place their gates on the plots! Flow data is messy, so learning how... will have numbers by gated populations; this indicates the percentage of total events contained in the...intracellular IFNγ, IL-17 and IL-10 producing T cells correlates with the occurrence of post-transplant opportunistic...

...instance, GFP may occlude catalytic sites or obstruct binding sites or motifs that are necessary for post-translational...avidity because it doubles the number of binding sites per unit.  There are several ways to measure stickiness...Where should the fluorescent protein be inserted? Sites that are less likely to disrupt the structure of...this is not conclusive evidence that the fusion operates in the same way as the native protein. Although...PMC4927352. Janetopoulos, Chris, Tian Jin, and Peter Devreotes. "Receptor-mediated activation of heterotrimeric...

...   This system is distinct from those used in eukaryotes in that CRISPR isn’t the primary editing force... similarly to CRISPR/Cas9 systems adapted for eukaryotes in that Cas9-mediated cleavage induces HDR to...design more familiar to those using CRISPR in eukaryotes (Qi et al., 2013). As in other systems, catalytically...either Gram-negative bacteria or Gram-positive firmicutes, to bring CRISPRi to a much wider swath of bacterial...Pseudomonas syringae. These vectors contain BsaI sites for easy cloning of a gRNA of interest. Cytidine...organisms in the genus. Work from the Sun lab demonstrates the utility of cytidine and adenine base editors...variant of the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig...

...follow #SfN14 on Twitter for live tweeting updates and the latest news from attendees. And for those of you...students and postdocs, which accounts for many, many lattes and espressos. The energy boost is well needed ...symposium discussing progress made in reprogramming astrocytes to functional neurons in vivo. And I’ve seen ...cup and get back to our booth, I will have more updates coming soon. Stop by booth #3431 if you’re at #...

...each tagged protein must be tested carefully to ensure it retains the attributes of the native protein. Since...accessibility to an antibody. From their initial pool of candidates, they selected 12 candidate protein domains from... from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed ...expressed well in cells and did not form undesirable aggregates. Using budding yeast as a model system, Georgieva...

...Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation rates than KTRs which is...import rates can influence KTR translocation. Work from the Covert lab suggests that these rates have negligible...negligible effect on KTRs, however nuclear translocation rates are another variable to consider when designing ...

... frequently used elements in plant expression cassettes: plant promoters, N-terminal tags, coding sequences...C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants. The complete... gives the option to stack several expression cassettes onto a single T-DNA in subsequent rounds, dramatically...always have the same recognition site but distinct sites where the enzyme will cut. You only need a single...available here.) Although Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle...

...efficiently transfer a range of protein sizes in minutes without any transfer buffer. These systems, however...Different immunoassays detect proteins in different states. For example, in immunoprecipitation (IP) the antibody...physiologically relevant levels. Some vendors will also test their antibody in a variety of tissues to demonstrate... methylation and ubiquitination. For example, to test if a protein is phosphorylated following a specific...for subcellular fractionations, a procedure that isolates proteins from different cellular compartments ...antibodies that are validated for your application and test the antibody in your specific system. If you found...

..."The greatest good you can do for another is not just to share your riches but to reveal to him his own...characteristics of a good mentor: Yoda image courtesy of jorenl/Vecteezy.com Helps set concrete... helps you reach your goals Listens well and communicates clearly and openly Gives honest feedback without...to take risks A great Mentor is often one that devotes time and energy to thinking about how to be a great...professor at a research university in the northeast, writes a thoughtful treatise on the kind of advisor/mentor...

...our in-stock viral preps: ddPCR, purity testing, sterility testing, and viral genomic sequencing to confirm...newsletter for updates on all things Addgene; or subscribe to the Addgene blog for updates and educational...that Addgene cannot perform functional or in vivo testing. We recommend you check the original paper of your...

...induce recombination at pseudo- or cryptic loxP sites (estimated to occur at a frequency of 1.2 per megabase...used successfully in Drosophila, so Rubin sought to test the suitability of four additional yeast recombinases...recombination site for each enzyme, were generated to test recombination efficiency. When expressed in neuronal...each recombinase and reporter construct pair were tested for excision efficiency. B2 and R showed partial...doi:10.1007/s004270100167. PubMed. Cryptic loxP sites in mammalian genomes: genome-wide distribution and...

...mismatched recognition sites, its presence induces first opsin inversion, and then lox sites excision, therefore...Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger neural...neural excitation, whereas yellow light activates halorhodopsins to silence neuronal activity (Figure 1) ...induce DNA recombination at defined recognition sites (i.e., loxP and FRT, respectively) (Abremski and...floxed”) by two sets of incompatible recognition sites (e.g., loxP and lox511) in opposite orientations...

...from the NIH, in the United States) Strong letters of reference that attest to your talent and expertise...fellowship lasts five years. Most universities and institutes annually advertise for Group Leader positions... a postdoctoral mentor. A good mentor typically writes the grants that fund your research, guides the ...Here is my personal list:   Exercise - nothing dissipates stress and improves your perspective like intense... Jennifer Lippincott-Schwartz at the National Institutes of Health, was an Associate Professor at Albert...