...field, let's look at how prokaryotes originally utilized CRISPR/Cas as a powerful and adaptive defense ...safeguard against bacteriophages did not come to light until 2007. Initial experiments exposed S. thermophilus...prokaryotes developed an adaptive immune system that utilized various cas genes not only to store a record of..., 83% of archaeal and 45% of bacterial genomes utilize this system (4). Makarova et al. grouped the systems... not translatable to the other CRISPR subtypes. Still, the large and dynamic range of CRISPR systems between...

... question of what I wanted to do when I grew up until I was all but finished my undergraduate degree (...of how to appeal to a given audience and how to utilize a wide array of media in doing so—can prove invaluable...of science communication, then, is its sheer versatility in the face of disparate spheres of research....incorporate other facets which occupied my attention, until these coalesced into a research topic on concussion...unsettled science. Understanding just how the public utilizes insights gleaned from scientific research is, ...

...generally agreed-upon international moratorium on utilizing CRISPR/Cas9 in humans. This topic was to be revisited...than having a meeting to agree upon a moratorium until another meeting, three branches – a private funder... unethical experimentation on human subjects, utilizing a scientific technique not proven adequately safe... of the consulting firm Rogue Bioethics. Kelly utilizes her firsthand experience in the software industry...all of whom are named Charlotte, all of whom are still working on theirpenmanship. Additional resources...

...extraction methods evolved into a simple protocol still used today There are many alternative methods for...-phenol-chloroform. In this method, samples are still homogenized and lysed in a guanidinium thiocyanate...hours, and variations on this no-kit method are still widely used today (Chomcynski and Sacchi, 2006)....Making the simple protocol even more foolproof (still without a kit!) As mentioned above, working with...with RNA requires keeping your samples cold until homogenization and cell lysis. This can be challenging...

...think I would have had the nerve to do what I did. Until that panicked, split-second decision, I had always...mind went blank, my options were to stop talking until I could present something coherent or to quickly...related to this paper and/or biology in general until I reached Fig. 6, Panel L, and hope nobody noticed... take that step. Repeat that as often as needed until you get to the end of the presentation, at which... to be anxious, nervous, and/or apprehensive. I still get nervous quite regularly, and I've had multiple...

... and following citations through the literature until you have all of the information you need. Reading..., but wait to sort out any of the finer details until you see the figures in the results section. As you...experiments are geared towards eliminating possibilities until the results focus in on a particular conclusion.... you prefer, printing and hand-writing notes is still possible, but not feasible for every paper you will...search. Writing this tutorial has shown me that I’m still in the dark ages with respect to my organization...

...CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability...Table 1). Additionally, hiPSCs and mouse BM cells still behaved like iPSCs and BM cells after Nanoblade ... like the addition of a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence... easily produced in a BSL2 lab, making them a versatile and accessible genome editing tool.   References...

...proteins that don’t utilize NGG   There are plenty of Cas9 variants which utilize alternative PAMs. These...how close is close enough for the desired edit to still occur if the cut site is sub-optimal? As with everything...et al, 2017). To add insult to injury, the most utilized PAM sequence, that of SpCas9, is NGG. Assuming...Nonetheless, in an AT-rich region, these Cas9s may still leave you choosing between efficiency and location...

... as “instillation” or “gavage,” as they do not require any punctures. Intratracheal instillation Intratracheal...Skill required: advanced Intranasal instillation Intranasal instillation is very similar to its intratracheal...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...

...after graduate school, at least not while you’re still in the thick of it. About three or four years into...ways to get a taste of a variety of careers while still working towards your degree. For me, experimenting... said, “Boy, I love science,” and I realized, I still do too! Beyond the outreach experience I gained ...back into lab the next day and remembered why I still love science. Keep building on your time in graduate...look back at a positive, fulfilling experience. I still use my network and graduate school resources daily...

...behind at the site of excision, this approach is still not considered safe for therapeutic applications...screening cells for the presence of transgenes is still an important control. Additionally, Sendai virus...The typical drawbacks of using transfection will still exist: transfection efficiency is cell-type dependent...by dilution with each cell division, but it can still take several passages for the minicircle to be completely... that ⅔ of subclones generated from EBNA1 iPSCs still had episome DNA present. DNA-Free Methods RNA...reprogramming proteins for 8 hours a week for 6 weeks, and still required further culturing to generate iPSC colonies...

...visualization techniques. In addition, their incredible versatility opens aptamers for use in many entirely new applications...signal-off” biosensors in which the aptamers fluoresce until they bind to the appropriate ligand, but relying...the bound protein fluorophores and quenchers are still relatively bulky. One of the advantages of oligonucleotides...stable and brighter version of the aptamer, but it still had limitations in vivo. Using a combination of ... a lower magnesium dependence. Though initially still unstable in eukaryotes, the Jaffrey lab has since...

...experiments. There are far too many behaviors that have utilized optogenetics to be fully summarized in a short...circuit is sufficient to induce reinforcement (instills a sense of “reward”), then the animal will repeatedly...type of behavior, certain technical considerations still apply. Testing stimulation First, In lieu of electrophysiology...prove inhibition of neuronal activation, but it may still be possible to use c-fos expression to get the evidence...advances of some are shared with all. Due to the versatility and standardization of genetic tools in the laboratory...

...Easy-to-use tools to monitor these features were lacking until Roberta Gottlieb and Zhen Yan labs developed MitoTimer...constructed using Golden GATEway cloning have been utilized to create recombination template vectors, to perform...cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly steps. Before beginning... (SSA) assay. This new Platinum Gate TALEN Kit utilizes a 4-module assembly system, which reduces the ... of synapses in the mouse brain. Kim et al is utilizing a mammalian GRASP (GFP reconstitution across synaptic... were reported in 2007. In order to extend the utility of the system, the researchers developed three ...plasmid kit page or read our blog post: Quick, Versatile Plant Transgenesis with GreenGate Plasmids. ...

...plasmid replaces the chromosomal copy. This is a versatile technique, and is less prone to off-target mutations...counter-selection system relies on the sacB gene product of B. subtilis (Hmelo et al., 2015). The sacB gene product is...that have only gone through the single cross over still contain sacB and will not grow on sucrose. One ...et al., 2014). Replaced the pDS132 MCS with a versatile, synthetic MCS (designed with the help of the ...and continue to improve the efficiency of this versatile technique. Many thanks to our guest blogger Jacob...

...or cell culture. For example, Scott Lowe’s lab utilized mice to study how different mutations in leukemia...at a time. Zebrafish embryos are also laid and fertilized externally, allowing scientists to easily manipulate...sizes. The majority of C. elegans are also self-fertile hermaphrodites making it easy to conduct large ...to name a few. For example, Andrew Leifer’s lab utilized the worm to study calcium signaling in the entire...PMC3703927. Jennings, Barbara H. "Drosophila–a versatile model in biology & medicine." Materials today ...

...other cellular proteins. BRET-based biosensors utilizing NanoLuc® luciferase Many intracellular sensors... activation of other fluorescent probes but can still transfer energy to the fluorescent protein acceptor...researchers to convert FRET biosensors into more versatile BRET biosensors that can even be paired with optogenetic... from Addgene and we're excited to see how you utilize them in your group! Many thanks to our guest blogger...Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chem ...

...nucleotide transitions but still requires a nearby PAM, although these are still early days for this technology...gRNAs to 5 – 65% of the protein coding region will still result in many dozens of gRNAs to choose from. With... While SpCas9, with a PAM preference of NGG, is still the most widely-used Cas enzyme, the development...basic landscape of mismatches that can nevertheless still lead to activity has been established, and can be...

...spectrum and emission spectrum. Fluorescent dyes utilize these fluorophores by combining them with a molecular... tags, such as higher specificity, increased versatility in probe attachment, and compatibility with far-red...These variations directly impact the way you can utilize these dyes. The most-used fluorescent dye is fluorescein...are currently offered at Promega. Due to the versatility of interchangeable labeling using the HaloTag...driven by the need for more precise, stable, and versatile imaging tools in biological research. This does...

...Tags. Beyond E. coli: purifying from Bacillus subtilis and Pichia pastoris pCri also includes plasmids...than E. coli, specifically the bacterium Bacillus subtilis and the yeast Pichia pastoris. The improved secretory.... pCri vector pCri-13a, designed for use in B. subtilis, contains the MISTIC (membrane-integrating sequence...proteins that natively localize there. pCri is both versatile and easy to use, and we hope that it will take...