Developing high-efficiency base editors by combining optimized synergistic core components with new types of nuclear localization signal peptide

The clustered regularly interspaced short palindromic repeats(CRISPR)–CRISPR-associated protein(Cas) system has been widely used for genome editing. In this system, the cytosine base editor(CBE) and adenine base editor(ABE) allow generating precise and irreversible base mutations in a programmable manner and have been used in many different types of cells and organisms. However, their applications are limited by low editing efficiency at certain genomic target sites or at specific target cytosine(C) or adenine(A) residues. Using a strategy of combining optimized synergistic core components, we developed a new multiplex super-assembled ABE(sABE) in rice that showed higher base-editing efficiency than previously developed ABEs. We also designed a new type of nuclear localization signal(NLS) comprising a FLAG epitope tag with four copies of a codon-optimized NLS(F4NLS^(r2)) to generate another ABE named F4NLS-sABE. This new NLS increased editing efficiency or edited additional A at several target sites. A new multiplex super-assembled CBE(sCBE) and F4NLS^(r2) involved F4NLS-sCBE were also created using the same strategy. F4NLS-sCBE was proven to be much more efficient than sCBE in rice. These optimized base editors will serve as powerful genome-editing tools for basic research or molecular breeding in rice and will provide a reference for the development of superior editing tools for other plants or animals.

A comparative study on crash-influencing factors by facility types on urban expressway

This study aims at identifying crash-influencing factors by facility type of Nagoya Urban Expressway, considering the interaction of geometry, traffic flow, and ambient conditions. Crash rate （CR） model is firstly developed separately at four facility types： basic, merge, and diverge segments and sharp curve. Traffic flows are thereby categorized, and based on the traffic categories, the significances of factors affecting crashes are analyzed by principal component analysis. The results reveal that, the CR at merge segment is significantly higher than those at basic and diverge segments in uncongested flow, while the value is not significantly different at the three facility types in congested flow. In both un- and congested flows, sharp curve has the worst safety performance in view of its highest CR. Regarding influencing factors, geometric design and traffic flow are most significant in un- and congested flows, respectively. As mainline flow increases, the effect of merging ratio affecting crash is on the rise at basic and merge segments as opposed to the decreasing significance of diverging ratio at diverge segment. Mean- while, longer acceleration and deceleration lanes are adverse to safety in uncongested flow, while shorter acceleration and deceleration lanes are adverse in congested flow. Due to its special geometric design, crashes at sharp curve are highly associated with the large centrifugal force and heavy restricted visibility.

Evaluation of COSMO-RS for solid–liquid equilibria prediction of binary eutectic solvent systems

For the design of eutectic solvents(ESs,usually also known as deep eutectic solvents),the prediction of the solid–liquid equilibria(SLE)between candidate components is of primary relevance.In the present work,the SLE prediction of binary eutectic solvent systems by the COSMO-RS model is systematically evaluated,thereby examining the applicability of this method for ES design.Experimental SLE of such systems are first collected exhaustively from the literature,following which COSMO-RS SLE calculations are accordingly carried out.By comparing the experimental and predicted eutectic points(eutectic temperature and eutectic composition)of the involved systems,the effects of salt component conformer and COSMO-RS parameterization as well as the applicability for different types of components(specifically the second component paired with the first salt one)are identified.The distinct performances of COSMO-RS SLE prediction for systems involving different types of components are further interpreted from the non-ideality and fusion enthalpy point of view.

Hyper-spectral characteristics and classification of farmland soil in northeast of China

The physical and chemical heterogeneities of soils make the soil spectral different and complicated, and it is valuable to increase the accuracy of prediction models for soil organic matter（SOM） based on pre-classification. This experiment was conducted under a controllable environment, and different soil samples from northeast of China were measured using ASD2500 hyperspectral instrument. The results showed that there are different reflectances in different soil types. There are statistically significant correlation between SOM and reflectence at 0.05 and 0.01 levels in 550–850 nm, and all soil types get significant at 0.01 level in 650–750 nm. The results indicated that soil types of the northeast can be divided into three categories： The first category shows relatively flat and low reflectance in the entire band; the second shows that the spectral reflectance curve raises fastest in 460–610 nm band, the sharp increase in the slope, but uneven slope changes; the third category slowly uplifts in the visible band, and its slope in the visible band is obviously higher than the first category. Except for the classification by curve shapes of reflectance, principal component analysis is one more effective method to classify soil types. The first principal component includes 62.13–97.19% of spectral information and it mainly relates to the information in 560–600, 630–690 and 690–760 nm. The second mainly represents spectral information in 1 640–1 740, 2 050–2 120 and 2 200–2 300 nm. The samples with high OM are often in the left, and the others with low OM are in the right of the scatter plot（the first principal component is the horizontal axis and the second is the longitudinal axis）. Soil types in northeast of China can be classified effectively by those two principles; it is also a valuable reference to other soil in other areas.