In vivo genome editing using Staphylococcus aureus Cas9

The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that employ the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we characterize six smaller Cas9 orthologs and show that Cas9 from Staphylococcus aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9, while being >1kb shorter. We packaged SaCas9 and its sgRNA expression cassette into a single AAV vector and targeted the cholesterol regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we observed >40% gene modification, accompanied by significant reductions in serum Pcsk9 and total cholesterol levels. We further demonstrate the power of using BLESS to assess the genome-wide targeting specificity of SaCas9 and SpCas9, and show that SaCas9 can mediate genome editing in vivo with high specificity

Sensing with nanopores--the influence of asymmetric blocking on electrochemical redox cycling current.

Nanoporous redox cycling devices are highly efficient tools for the electrochemical sensing of redox-active molecules. By using a redox-active mediator, this concept can be exploited for the detection of molecular binding events via blocking of the redox cycling current within the nanopores. Here, we investigate the influence of different blocking scenarios inside a nanopore on the resulting redox cycling current. Our analysis is based on random walk simulations and finite element calculations. We distinguish between symmetric and asymmetric pore blocking and show that the current decrease is more pronounced in the case of asymmetric blocking reflecting the diffusion-driven pathway of the redox-active molecules. Using random walk simulations, we further study the impact of pore blocking in the frequency domain and identify relevant features of the power spectral density, which are of particular interest for sensing applications based on fluctuation analysis

The influence of supporting ions on the electrochemical detection of individual silver nanoparticles: Understanding the shape and frequency of current transients in nano-impacts.

We report the influence of electrolyte composition and concentration on the stochastic amperometric detection of individual silver nanoparticles at microelectrode arrays and show that the sensor response at certain electrode potentials is dependent on both the conductivity of the electrolyte and the concentration of chloride ions. We further demonstrate that the chloride concentration in solution heavily influences the characteristic current spike shape of recorded nanoparticle impacts: While typically too short to be resolved in the measured current, the spike widths are significantly broadened at low chloride concentrations below 10 mM and range into the millisecond regime. The analysis of more than 25.000 spikes reveals that this effect can be explained by the diffusive mass transport of chloride ions to the nanoparticle, which limits the oxidization rate of individual silver nanoparticles to silver chloride at the chosen electrode potential

Sex-related differences and age of peak performance in breaststroke versus freestyle swimming

BACKGROUND: Sex-related differences in performance and in age of peak performance have been reported for freestyle swimming. However, little is known about the sex-related differences in other swimming styles. The aim of the present study was to compare performance and age of peak performance for elite men and women swimmers in breaststroke versus freestyle. METHODS: Race results were analyzed for swimmers at national ranked in the Swiss high score list (during 2006 through 2010) and for international swimmers who qualified for the finals of the FINA World Swimming Championships (during 2003 through 2011). RESULTS: The sex-related difference in swimming speed was significantly greater for freestyle than for breaststroke over 50 m, 100 m, and 200 m race distances for Swiss swimmers, but not for FINA finalists. The sex-related difference for both freestyle and breaststroke swimming speeds decreased significantly with increasing swimming distance for both groups. Race distance did not affect the age of peak performance by women in breaststroke, but age of peak performance was four years older for FINA women than for Swiss women. Men achieved peak swimming performance in breaststroke at younger ages for longer race distances, and the age of peak swimming performance was six years older for FINA men than for Swiss men. In freestyle swimming, race distance did not affect the age of peak swimming performance for Swiss women, but the age of peak swimming performance decreased with increasing race distance for Swiss men and for both sexes at the FINA World Championships. CONCLUSIONS: Results of the present study indicate that (i) sex-related differences in swimming speed were greater for freestyle than for breaststroke for swimmers at national level, but not for swimmers at international level, and (ii) both female and male swimmers achieved peak swimming speeds at younger ages in breaststroke than in freestyle. Further studies are required to better understand differences between trends at national and international levels

Measurements of direct drive laser imprint in thin foils by radiography using an x-ray laser backlighter

In direct drive inertial confinement fusion, the residual speckle pattern remaining after beam smoothing plays an important role in the seeding of instabilities at the ablation front. An x-ray laser is used as an extreme ultraviolet backlighter to characterize the imprinted modulation in thin foils for smoothing by random phase plate and by spectral dispersion for both 0.35 and 0.53 mu m irradiation, and by induced spatial incoherence for 0.53 mu m irradiation. Measurements of the imprinted modulation due to a single optical mode generated by two beam interference, and modification of the imprint with a superposed smooth irradiation to study time dependence of the imprinting process are demonstrated. (C) 1997 American Institute of Physics