Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants in Brassica.

CRISPR/Cas9, presently the most widely used genome editing technology, has provided great potential for functional studies and plant breeding. However, the strict requirement for a protospacer adjacent motif (PAM) has hindered the application of the CRISPR/Cas9 system because the number of targetable genomic sites is limited. Recently, the engineered variants Cas9-NG, SpG, and SpRY, which recognize non-canonical PAMs, have been successfully tested in plants (mainly in rice, a monocot). In this study, we evaluated the targeted mutagenesis capabilities of these Cas9 variants in two important Brassica vegetables, Chinese cabbage (Brassica rapa spp. pekinensis) and cabbage (Brassica oleracea var. capitata). Both Cas9-NG and SpG induced efficient mutagenesis at NGN PAMs, while SpG outperformed Cas9-NG at NGC and NGT PAMs. SpRY achieved efficient editing at almost all PAMs (NRN > NYN), albeit with some self-targeting activity at transfer (T)-DNA sequences. And SpRY-induced mutants were detected in cabbage plants in a PAM-less fashion. Moreover, an adenine base editor was developed using SpRY and TadA8e deaminase that induced A-to-G conversions within target sites using non-canonical PAMs. Together, the toolboxes developed here induced successful genome editing in Chinese cabbage and cabbage. Our work further expands the targeting scope of genome editing and paves the way for future basic research and genetic improvement in Brassica. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-024-00155-7.

Magnetic field modulation effect of photoelectric properties in dye-sensitized solar cells with La0.67(Ca,Ba)0.33MnO3 as counter electrodes.

BACKGROUND: In recent years, many semiconductor materials with unique band structures have been used as Pt counter electrode (CE) substitutes for dye-sensitized solar cells (DSSCs), which makes the photoelectric properties of DSSCs possible to be modulated by electric field, magnetic field, and light field. In this work, La0.67(Ca Ba)0.33MnO3 (LCBMO) thin film is employed to act as CE in DSSCs. METHOD: The experimental results indicate that short-circuit current density and photoelectric conversion efficiency present better stability when applying an external magnetic field to the DSSCs. Furthermore, both the exchange current density (J0) and limit diffusion current density (Jlim) are largely enhanced by an external magnetic field. J0 increases from -0.51 mA•cm-2 to -0.65 mA•cm-2, and Jlim increases from 0.2 mA•cm-2 to 0.3 mA•cm-2 when applying a magnetic field of 0.25 T. RESULT: The fitting results of the impedance test verify that the magnetic field reduces the value of Rct. CONCLUSION: Both magnetic-field enhancing catalytic activity and CMR effect jointly promote the increase of photocurrent and finally improve the photovoltaic effect in DSSCs.

Prime editing: Its systematic optimization and current applications in disease treatment and agricultural breeding.

CRISPR/Cas-mediated genome-editing technology has accelerated the development of the life sciences. Prime editing has raised genome editing to a new level because it allows for all 12 types of base substitutions, targeted insertions and deletions, large DNA fragment integration, and even combinations of these edits without generating DNA double-strand breaks. This versatile and game-changing technology has successfully been applied to human cells and plants, and it currently plays important roles in basic research, gene therapy, and crop breeding. Although prime editing has substantially expanded the range of possibilities for genome editing, its efficiency requires improvement. In this review, we briefly introduce prime editing and highlight recent optimizations that have improved the efficiency of prime editors. We also describe how the dual-pegRNA strategy has expanded current editing capabilities, and we summarize the potential of prime editing in treating mammalian diseases and improving crop breeding. Finally, we discuss the limitations of current prime editors and future prospects for optimizing these editors.