Laboratory Exercises Caenorhabditis elegans as an Undergraduate Educational Tool for Teaching RNAi*

Discovery of RNA-mediated interference (RNAi) is widely recognized as one of the most significant molecular biology breakthroughs in the past 10 years. There is a need for science educators to develop teaching tools and laboratory activities that demonstrate the power of this new technology and help students to better understand the RNAi process. C. elegans is an ideal model organism for the undergraduate laboratory because of the simplicity of worm maintenance, its well-studied genetic background, and the fact that it can be employed as a model organism in laboratory environments where vertebrate research is restricted. Certain unique features of C. elegans make it a very suitable organism for RNAi studies. Specifically, nematode strains highly sensitive to RNAi are readily available from public sources, and RNAi induction by a feeding method is an uncomplicated procedure that lends itself readily as an educational tool. In this article, we provide a detailed depiction of the use of C. elegans as an RNAi educational tool, describing two separate RNAi-based experiments. One is a qualitative experiment where students can examine the effects of knocking down the unc-22 gene involved in the regulation of muscle contraction, which results in a ''twitching'' phenotype. The other experiment is a quantitative RNAi experiment, where students measure the effect of knocking down the lsy-2 gene involved in neuronal development. Although these experiments are designed for a college-level study, nematode research projects can also be accomplished in secondary school facilities

All-dielectric magnetic metasurface for advanced light control in dual polarizations combined with high-Q resonances

Nanostructured magnetic materials provide an efficient tool for light manipulation on sub-nanosecond and sub-micron scales, and allow for the observation of the novel effects which are fundamentally impossible in smooth films. For many cases of practical importance, it is vital to observe the magneto-optical intensity modulation in a dual-polarization regime. However, the nanostructures reported on up to date usually utilize a transverse Kerr effect and thus provide light modulation only for p-polarized light. We present a concept of a transparent magnetic metasurface to solve this problem, and demonstrate a novel mechanism for magneto-optical modulation. A 2D array of bismuth-substituted iron-garnet nanopillars on an ultrathin iron-garnet slab forms a metasurface supporting quasi-waveguide mode excitation. In contrast to plasmonic structures, the all-dielectric magnetic metasurface is shown to exhibit much higher transparency and superior quality-factor resonances, followed by a multifold increase in light intensity modulation. The existence of a wide variety of excited mode types allows for advanced light control: transmittance of both p- and s-polarized illumination becomes sensitive to the medium magnetization, something that is fundamentally impossible in smooth magnetic films. The proposed metasurface is very promising for sensing, magnetometry and light modulation applications

Optical and thermal analysis of the light-heat conversion process employing an antenna-based hybrid plasmonic waveguide for HAMR

We investigate a tapered, hybrid plasmonic waveguide which has previously been proposed as an optically efficient near-field transducer (NFT), or component thereof, in several devices which aim to exploit nanofocused light. We numerically analyze how light is transported through the waveguide and ultimately focused via effective-mode coupling and taper optimization. Crucial dimensional parameters in this optimization process are identified that are not only necessary to achieve maximum optical throughput, but also optimum thermal performance with specific application towards heat-assisted magnetic recording (HAMR). It is shown that existing devices constructed on similar waveguides may benefit from a heat spreader to avoid deformation of the plasmonic element which we achieve with no cost to the optical efficiency. For HAMR, our design is able to surpass many industry requirements in regard to both optical and thermal efficiency using pertinent figure of merits like 8.5% optical efficiency.Comment: 14 pages, 7 figures, and 3 tables. Published version: see https://doi.org/10.1364/OE.26.001752. Related works: see https://doi.org/10.1364/oe.22.011236, https://doi.org/10.1364/oe.26.030292, and https://doi.org/10.1063/5.0044490. Keywords: Integrated Optics; Components; Integrated Optics Devices; Surface Plasmons; Plasmonic

Optical and thermal analysis of the light-heat conversion process employing an antenna-based hybrid plasmonic waveguide for HAMR

We investigate a tapered, hybrid plasmonic waveguide which has previously been proposed as an optically efficient near-field transducer (NFT), or component thereof, in several devices which aim to exploit nanofocused light. We numerically analyze how light is transported through the waveguide and ultimately focused via effective-mode coupling and taper optimization. Crucial dimensional parameters in this optimization process are identified that are not only necessary to achieve maximum optical throughput, but also optimum thermal performance with specific application towards heat-assisted magnetic recording (HAMR). It is shown that existing devices constructed on similar waveguides may benefit from a heat spreader to avoid deformation of the plasmonic element which we achieve with no cost to the optical efficiency. For HAMR, our design is able to surpass many industry requirements in regard to both optical and thermal efficiency using pertinent figure of merits like 8.5% optical efficiency

Optical, thermal, and bit-writing analysis of a directly coupled plasmonic waveguide for heat-assisted magnetic recording

We investigate the energy conversion process and subsequent thermal and bit-writing performance of a plasmonic near-field transducer (NFT) under steady-state operation within heat-assisted magnetic recording (HAMR) devices. The NFT is composed of metal-insulator-metal (MIM) layers that are designed to localize heating and produce optimal thermal gradients in order to relieve parasitic heating effects in the NFT. The thin-film MIM structure confines the electromagnetic energy in the down-track direction while cross-track confinement is achieved by tapering the insulator feature of the MIM. A comparative analysis using Gold and a number of novel Au alloys is undertaken. Modeled performance shows excellent thermal spot confinement (50 × 50 nm2) of temperatures above 650 K at an input laser power of 830 nm of less than 5 milliwatts. In addition, micromagnetic simulations using a stochastic Landau-Lifshitz-Bloch equation yield excellent signal to noise ratio with minimum jitter of under 2 nm when recording

Autoluminescent Plants

Prospects of obtaining plants glowing in the dark have captivated the imagination of scientists and layman alike. While light emission has been developed into a useful marker of gene expression, bioluminescence in plants remained dependent on externally supplied substrate. Evolutionary conservation of the prokaryotic gene expression machinery enabled expression of the six genes of the lux operon in chloroplasts yielding plants that are capable of autonomous light emission. This work demonstrates that complex metabolic pathways of prokaryotes can be reconstructed and function in plant chloroplasts and that transplastomic plants can emit light that is visible by naked eye

Profiling cytotoxic microRNAs in pediatric and adult glioblastoma cells by high-content screening, identification, and validation of miR-1300

MicroRNAs play an important role in the regulation of mRNA translation and have therapeutic potential in cancer and other diseases. To profile the landscape of microRNAs with significant cytotoxicity in the context of glioblastoma (GBM), we performed a high-throughput screen in adult and pediatric GBM cells using a synthetic oligonucleotide library representing all known human microRNAs. Bioinformatics analysis was used to refine this list and the top seven microRNAs were validated in a larger panel of GBM cells using state-of-the-art in vitro assays. The cytotoxic effect of our most relevant candidate was assessed in a preclinical model. Our screen identified ~100 significantly cytotoxic microRNAs with 70% concordance between cell lines. MicroRNA-1300 (miR-1300) was the most potent and robust candidate. We observed a striking binucleated phenotype in miR-1300 transfected cells due to cytokinesis failure followed by apoptosis. This was also observed in two stem-like patient-derived cultures. We identified the physiological role of miR-1300 as a regulator of endomitosis in megakaryocyte differentiation where blockade of cytokinesis is an essential step. In GBM cells, where miR-1300 is normally not expressed, the oncogene Epithelial Cell Transforming 2 (ECT2) was validated as a direct key target. ECT2 siRNA phenocopied the effects of miR-1300, and ECT2 overexpression led to rescue of miR-1300 induced binucleation. We showed that ectopic expression of miR-1300 led to decreased tumor growth in an orthotopic GBM model. Our screen provides a resource for the neuro-oncology community and identified miR-1300 as a novel regulator of endomitosis with translatable potential for therapeutic application