Germline Cas9 expression yields highly efficient genome engineering in a major worldwide disease vector, Aedes aegypti.

The development of CRISPR/Cas9 technologies has dramatically increased the accessibility and efficiency of genome editing in many organisms. In general, in vivo germline expression of Cas9 results in substantially higher activity than embryonic injection. However, no transgenic lines expressing Cas9 have been developed for the major mosquito disease vector Aedes aegypti Here, we describe the generation of multiple stable, transgenic Ae. aegypti strains expressing Cas9 in the germline, resulting in dramatic improvements in both the consistency and efficiency of genome modifications using CRISPR. Using these strains, we disrupted numerous genes important for normal morphological development, and even generated triple mutants from a single injection. We have also managed to increase the rates of homology-directed repair by more than an order of magnitude. Given the exceptional mutagenic efficiency and specificity of the Cas9 strains we engineered, they can be used for high-throughput reverse genetic screens to help functionally annotate the Ae. aegypti genome. Additionally, these strains represent a step toward the development of novel population control technologies targeting Ae. aegypti that rely on Cas9-based gene drives

Attention is more than prediction precision [Commentary on target article]

A cornerstone of the target article is that, in a predictive coding framework, attention can be modelled by weighting prediction error with a measure of precision. We argue that this is not a complete explanation, especially in the light of ERP (event-related potentials) data showing large evoked responses for frequently presented target stimuli, which thus are predicted

Arbitrary multi-site two-photon excitation in four dimensions

We demonstrate dynamic and arbitrary multisite two-photon excitation in three dimensions using the holographic projection method. Rapid response (fourth dimension) is achieved through high-speed noniterative calculation of the hologram using a video graphics accelerator board. We verify that the projected asymmetric spot configurations have sufficient spatiotemporal photon density for localized two-photon excitation. This system is a significant advance and can be applied to time-resolved photolysis of caged compounds in biological cells and complex neuronal networks, nonlinear microfabrication and volume holographic optical storage.Comment: 10 pages including 4 figure

Characterizing the ZrBe2Hx Phase Diagram via Neutron Scattering Methods

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White Matter Tract Covariance Patterns Predict Age-Declining Cognitive Abilities

Previous studies investigating the relationship of white matter (WM) integrity to cognitive abilities and aging have either focused on a global measure or a few selected WM tracts. Ideally, contribution from all of the WM tracts should be evaluated at the same time. However, the high collinearity among WM tracts precludes systematic examination of WM tracts simultaneously without sacrificing statistical power due to stringent multiple-comparison corrections. Multivariate covariance techniques enable comprehensive simultaneous examination of all WM tracts without being penalized for high collinearity among observations. METHOD: In this study, Scaled Subprofile Modeling (SSM) was applied to the mean integrity of 18 major WM tracts to extract covariance patterns that optimally predicted four cognitive abilities (perceptual speed, episodic memory, fluid reasoning, and vocabulary) in 346 participants across ages 20 to 79years old. Using expression of the covariance patterns, age-independent effects of white matter integrity on cognition and the indirect effect of WM integrity on age-related differences in cognition were tested separately, but inferences from the indirect analyses were cautiously made given that cross-sectional data set was used in the analysis. RESULTS: A separate covariance pattern was identified that significantly predicted each cognitive ability after controlling for age except for vocabulary, but the age by WM covariance pattern interaction was not significant for any of the three abilities. Furthermore, each of the patterns mediated the effect of age on the respective cognitive ability. A distinct set of WM tracts was most influential in each of the three patterns. The WM covariance pattern accounting for fluid reasoning showed the most number of influential WM tracts whereas the episodic memory pattern showed the least number. CONCLUSION: Specific patterns of WM tracts make significant contributions to the age-related differences in perceptual speed, episodic memory, and fluid reasoning but not vocabulary. Other measures of brain health will need to be explored to reveal the major influences on the vocabulary ability

Evaluation and attribution of OCO-2 XCO_2 uncertainties

Evaluating and attributing uncertainties in total column atmospheric CO_2 measurements (XCO_2) from the OCO-2 instrument is critical for testing hypotheses related to the underlying processes controlling XCO_2 and for developing quality flags needed to choose those measurements that are usable for carbon cycle science. Here we test the reported uncertainties of version 7 OCO-2 XCO_2 measurements by examining variations of the XCO_2 measurements and their calculated uncertainties within small regions (∼  100 km  ×  10.5 km) in which natural CO_2 variability is expected to be small relative to variations imparted by noise or interferences. Over 39 000 of these small neighborhoods comprised of approximately 190 observations per neighborhood are used for this analysis. We find that a typical ocean measurement has a precision and accuracy of 0.35 and 0.24 ppm respectively for calculated precisions larger than  ∼  0.25 ppm. These values are approximately consistent with the calculated errors of 0.33 and 0.14 ppm for the noise and interference error, assuming that the accuracy is bounded by the calculated interference error. The actual precision for ocean data becomes worse as the signal-to-noise increases or the calculated precision decreases below 0.25 ppm for reasons that are not well understood. A typical land measurement, both nadir and glint, is found to have a precision and accuracy of approximately 0.75 and 0.65 ppm respectively as compared to the calculated precision and accuracy of approximately 0.36 and 0.2 ppm. The differences in accuracy between ocean and land suggests that the accuracy of XCO2 data is likely related to interferences such as aerosols or surface albedo as they vary less over ocean than land. The accuracy as derived here is also likely a lower bound as it does not account for possible systematic biases between the regions used in this analysis

Leading-order calculation of hadronic contributions to the muon g−2g-2 using the Dyson-Schwinger approach

We present a calculation of the hadronic vacuum polarization (HVP) tensor within the framework of Dyson--Schwinger equations. To this end we use a well-established phenomenological model for the quark-gluon interaction with parameters fixed to reproduce hadronic observables. From the HVP tensor we compute both the Adler function and the HVP contribution to the anomalous magnetic moment of the muon, $a_\mu$. We find $a_\mu^{HVP}= 6760\times 10^{-11}$ which deviates about two percent from the value extracted from experiment. Additionally, we make comparison with a recent lattice determination of $a_\mu^{HVP}$ and find good agreement within our approach. We also discuss the implications of our result for a corresponding calculation of the hadronic light-by-light scattering contribution to $a_\mu$.Comment: 9 pages, 8 figure

Functional and Biochemical Characterization of Dib1\u27s Role in Pre-Messenger RNA Splicing

The spliceosome is a dynamic macromolecular machine that undergoes a series of conformational rearrangements as it transitions between the several states required for accurate splicing. The transition from the B to Bact is a key part of spliceosome assembly and is defined by the departure of several proteins, including essential U5 component Dib1. Recent structural studies suggest that Dib1 has a role in preventing premature spliceosome activation, as it is positioned adjacent to the U6 snRNA ACAGAGA and the U5 loop I, but its mechanism is unknown. Our data indicate that Dib1 is a robust protein that tolerates incorporation of many mutations, even at positions thought to be key for its folding stability. However, we have identified two temperature-sensitive mutants that stall in vitro splicing prior to the first catalytic step and block assembly at the B complex. In addition, Dib1 readily exchanges in splicing extracts despite being a central component of the U5 snRNP, suggesting that the binding site of Dib1 is flexible. Structural analyses show that the overall conformation of Dib1 and the mutants are not affected by temperature, so the temperature sensitive defects most likely result from altered interactions between Dib1 and other spliceosomal components. Together, these data lead to a new understanding of Dib1\u27s role in the B to Bact transition and provide a model for how dynamic protein–RNA interactions contribute to the correct assembly of a complex molecular machine