Two recently engineered SpCas9 variants, namely xCas9 and Cas9-NG, show promising potential in improving targeting specificity and broadening the targeting range. In this study, we evaluated these Cas9 variants in the model and crop plant, rice. We first tested xCas9-3.7, the most effective xCas9 variant in mammalian cells, for targeted mutagenesis at 16 possible NGN PAM (protospacer adjacent motif) combinations in duplicates. xCas9 exhibited nearly equivalent editing efficiency to wild-type Cas9 (Cas9-WT) at most canonical NGG PAM sites tested, whereas it showed limited activity at non-canonical NGH (H = A, C, T) PAM sites. High editing efficiency of xCas9 at NGG PAMs was further demonstrated with C to T base editing by both rAPOBEC1 and PmCDA1 cytidine deaminases. With mismatched sgRNAs, we found that xCas9 had improved targeting specificity over the Cas9-WT. Furthermore, we tested two Cas9-NG variants, Cas9-NGv1 and Cas9-NG, for targeting NGN PAMs. Both Cas9-NG variants showed higher editing efficiency at most non-canonical NG PAM sites tested, and enabled much more efficient editing than xCas9 at AT-rich PAM sites such as GAT, GAA, and CAA. Nevertheless, we found that Cas9-NG variants showed significant reduced activity at the canonical NGG PAM sites. In stable transgenic rice lines, we demonstrated that Cas9-NG had much higher editing efficiency than Cas9-NGv1 and xCas9 at NG PAM sites. To expand the base-editing scope, we developed an efficient C to T base-editing system by making fusion of Cas9-NG nickase (D10A version), PmCDA1, and UGI. Taken together, our work benchmarked xCas9 as a high-fidelity nuclease for targeting canonical NGG PAMs and Cas9-NG as a preferred variant for targeting relaxed PAMs for plant genome editing.

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In this paper, we present an example-based colorization technique robust to illumination differences between grayscale target and color reference images. To achieve this goal, our method performs color transfer in an illumination-independent domain that is relatively free of shadows and highlights. It first recovers an illumination-independent intrinsic reflectance image of the target scene from multiple color references obtained by web search. The reference images from the web search may be taken from different vantage points, under different illumination conditions, and with different cameras. Grayscale versions of these reference images are then used in decomposing the grayscale target image into its intrinsic reflectance and illumination components. We transfer color from the color reflectance image to the grayscale reflectance image, and obtain the final result by relighting with the illumination component of the target image. We demonstrate via several examples that our method generates results with excellent color consistency.

We investigate superfluidity, and the mechanism for creation of quantized vortices, in the relativistic regime. The general framework is a nonlinear Klein-Gordon equation in curved spacetime for a complex scalar field, whose phase dynamics gives rise to superfluidity. The mechanisms discussed are local inertial forces (Coriolis and centrifugal), and current-current interaction with an external source. The primary application is to cosmology, but we also discuss the reduction to the nonrelativistic nonlinear Schrödinger equation, which is widely used in describing superfluidity and vorticity in liquid helium and cold-trapped atomic gases.

Commercial 3D scene acquisition systems such as the Microsoft Kinect sensor can reduce the cost barrier of realizing mid-air interaction. However, since it can only sense hand position but not hand orientation robustly, current mid-air interaction methods for 3D virtual object manipulation often require contextual and mode switching to perform translation, rotation, and scaling, thus preventing natural continuous gestural interactions. A novel handle bar metaphor is proposed as an effective visual control metaphor between the user's hand gestures and the corresponding virtual object manipulation operations. It mimics a familiar situation of handling objects that are skewered with a bimanual handle bar. The use of relative 3D motion of the two hands to design the mid-air interaction allows us to provide precise controllability despite the Kinect sensor's low image resolution. A comprehensive repertoire of 3D manipulation operations is proposed to manipulate single objects, perform fast constrained rotation, and pack/align multiple objects along a line. Three user studies were devised to demonstrate the efficacy and intuitiveness of the proposed interaction techniques on different virtual manipulation scenarios.