A number of mitochondrial diseases in humans are caused by point mutations that could be corrected by base editors, but delivery of CRISPR guide RNAs into the mitochondria is difficult. In this study, we present mitochondrial DNA base editors (mitoBEs), which combine a transcription activator-like effector (TALE)-fused nickase and a deaminase for precise base editing in mitochondrial DNA. Combining mitochondria-localized, programmable TALE binding proteins with the nickase MutH or Nt.BspD6I(C) and either the single-stranded DNA-specific adenine deaminase TadA8e or the cytosine deaminase ABOBEC1 and UGI, we achieve A-to-G or C-to-T base editing with up to 77% efficiency and high specificity. We find that mitoBEs are DNA strand-selective mitochondrial base editors, with editing results more likely to be retained on the nonnicked DNA strand. Furthermore, we correct pathogenic mitochondrial DNA mutations in patient-derived cells by delivering mitoBEs encoded in circular RNAs. mitoBEs offer a precise, efficient DNA editing tool with broad applicability for therapy in mitochondrial genetic diseases.

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Based on the classification of automated driving by the SAE (Society of Automotive Engineers) and the working principle of the ECPBS (Electronically Controlled Pneumatic Brake system), the requirements and the control modes of the APRV (Automatic Pressure Regulating Valve) were concluded. Four structural configurations for APRV were proposed to meet the requirements of the ECPBS. To study the pressure regulating characteristics of the APRV of different structure configurations, a simulation model was established, and a test bench was built. Through experiments, the correctness and the reliability of the simulation model were verified. The pressure regulation characteristics of the APRV of different structure configurations under different control conditions were revealed, and the suitable levels in the SAE automated driving classifications for automatic pressure regulators of different structure configurations were determined; thus, the theoretical underpinning to improve driving safety and develop automated driving was provided.

In order to adapt the development of vehicle driving automation technology for driving conditions under different levels of automation and based on the independently invented LF automatic pressure regulating valve (LF-APRV) for electronically controlled pneumatic brake systems (ECPBS), the dynamic PWM coupling pressure regulation method is proposed. This method realizes pressure regulation by adjusting the duty cycle of the control signal of the LF-APRV at different stages in the pressure regulation cycle. A co-simulation model was established to verify the feasibility of the method, and a test system was built to verify the correctness of the co-simulation model. Through the test, the pressure regulation performance of dynamic PWM coupling pressure regulation method and conventional on/off pressure regulation method was compared. The results show that the new method can improve the stability of pressure regulation, although the response time increases; under the new method, the overshoot of the pressure rising from 0 to 0.5 MPa was reduced by 69%, and the overshoot of the pressure decreasing from 0.5 MPa to 0.2 MPa was basically 0. Finally, tests and simulations showed that the dynamic PWM coupling pressure regulation method can meet the continuous graded braking requirements of vehicles, and the pressure response has good tracking performance on the target pressure.