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A homing rescue gene drive with multiplexed gRNAs reaches high frequency in cage populations but generates functional resistance

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Journal of Genetics and Genomics 2024年第8期51卷 836-843页

作者： Shibo Hou Jingheng Chen Ruobing Feng Xuejiao Xu Nan Liang Jackson Champer center for Bioinformatics Center for Life SciencesSchool of Life SciencesPeking UniversityBeijing 100871China

CRISPR homing gene drives have considerable potential for managing populations of medically and agriculturally significant *** operate by Cas9 cleavage followed by homology-directed repair,copying the drive allele to the wild-type chromosome and thus increasing in frequency and spreading throughout a ***,resistance alleles formed by end-joining repair pose a significant *** address this,we create a homing drive targeting the essential hairy gene in Drosophila *** resistance alleles are recessive lethal,while drive carriers have a recoded“rescue”version of *** drive inheritance rate is moderate,and multigenerational cage studies show drive spread to 96%–97%of the ***,the drive does not reach 100%due to the formation of functional resistance alleles despite using four *** alleles have a large deletion but likely utilize an alternate start ***,revised designs targeting more essential regions of a gene may be necessary to avoid such functional *** of the rescue element’s native 3'UTR with a homolog from another species increases drive inheritance by 13%–24%.This was possibly because of reduced homology between the rescue element and surrounding genomic DNA,which could also be an important design consideration for rescue gene drives.

关键词： Genedrive Homing drive Rescueelement Population modification Multiplexed gRNAs Resistance alleles Cage study

Light-stabilized GIL1 suppresses PIN3 activity to inhibit hypocotyl gravitropism

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Journal of Integrative Plant Biology 2024年第9期66卷 1886-1897页

作者： Xiaolian Wang Yanfang Yuan Laurence Charrier Zhaoguo Deng Markus Geisler Xing Wang Deng Haodong Chen center for Plant Biology School of Life SciencesTsinghua UniversityBeijing 100084China Tsinghua‐Peking center for life sciences Beijing 100084China school of Advanced Agricultural sciences and school of life sciences Peking‐Tsinghua Center for Life SciencesPeking UniversityBeijing 100871China Department of Biology University of FribourgFribourg 1700Switzerland

Light and gravity coordinately regulate the directional growth of *** Gravitropic in the Light 1(GIL1)inhibits the negative gravitropism of hypocotyls in red and far-red light,but the underlying molecular mechanisms remain *** study found that GIL1 is a plasma membrane-localized *** endodermal cells of the upper part of hypocotyls,GIL1 controls the negative gravitropism of ***1 directly interacts with PIN3 and inhibits the auxin transport activity of *** of PIN3 suppresses the abnormal gravitropic response of gil1 *** GIL1 protein is unstable in darkness but it is stabilized by red and far-red ***,our data suggest that light-stabilized GIL1 inhibits the negative gravitropism of hypocotyls by suppressing the activity of the auxin transporter PIN3,thereby enhancing the emergence of young seedlings from the soil.

关键词： GIL1 gravitropism hypocotyls light PIN3

Ultrafast micro/nano-manufacturing of metastable materials for energy

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National Science Review 2024年第4期11卷 283-298页

作者： Xiaoya Cui Yanchang Liu Yanan Chen school of Materials Science and Engineering Key Laboratory of Advanced Ceramics and Machining Technology(Ministry of Education) and Tianjin Key Laboratory of Composite and Functional MaterialsTianjin University Ministry of Education Key Laboratory of protein sciences School of Life SciencesTsinghua University

The structural engineering of metastable nanomaterials with abundant defects has attracted much attention in energy-related fields. The high-temperature shock(HTS) technique, as a rapidly developing and advanced synthesis strategy, offers significant potential for the rational design and fabrication of high-quality nanocatalysts in an ultrafast, scalable, controllable and eco-friendly way. In this review, we provide an overview of various metastable micro-and nanomaterials synthesized via HTS, including single metallic and bimetallic nanostructures, high entropy alloys, metal compounds(e.g. metal oxides) and carbon nanomaterials. Note that HTS provides a new research dimension for nanostructures, i.e. kinetic modulation. Furthermore, we summarize the application of HTS—as supporting films for transmission electron microscopy grids—in the structural engineering of 2D materials, which is vital for the direct imaging of metastable materials. Finally, we discuss the potential future applications of high-throughput and liquid-phase HTS strategies for non-equilibrium micro/nano-manufacturing beyond energy-related *** is believed that this emerging research field will bring new opportunities to the development of nanoscience and nanotechnology in both fundamental and practical aspects.

关键词： high-temperature shock technique defect engineering metastable phase kinetic modulation non-equilibrium micro/nano-manufacturing

Subtraction of liposome signals in cryo-EM structural determination of protein-liposome complexes

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Chinese Physics B 2024年第8期33卷 569-577页

作者：李首卿李明王玉梅李雪明 Institute of Physics Chinese Academy of SciencesBeijing 100190China University of Chinese Academy of sciences Beijing 100049China Key Laboratory for protein sciences of Ministry of Education School of Life SciencesTsinghua UniversityBeijing 100084China State Key Laboratory of Membrane Biology School of Life SciencesTsinghua UniversityBeijing 100084China Tsinghua–Peking Joint center for life sciences Beijing 100084China Beijing Frontier Research center for Biological Structure Beijing 100084China school of life sciences Tsinghua UniversityBeijing 100084China Beijing Branch of Songshan Lake Materials Laboratory Beijing 100190China

Reconstituting membrane proteins in liposomes and determining their structure is a common method for determining membrane protein structures using single-particle cryo-electron microscopy(cryo-EM).However,the strong signal of liposomes under cryo-EM imaging conditions often interferes with the structural determination of the embedded membrane ***,we propose a liposome signal subtraction method based on single-particle two-dimensional(2D)classification average images,aimed at enhancing the reconstruction resolution of membrane *** analyzed the signal distribution characteristics of liposomes and proteins within the 2D classification average images of protein–liposome complexes in the frequency *** on this analysis,we designed a method to subtract the liposome signals from the original particle *** the subtraction,the accuracy of single-particle three-dimensional(3D)alignment was improved,enhancing the resolution of the final 3D *** demonstrated this method using a PIEZO1-proteoliposome dataset by improving the resolution of the PIEZO1 protein.

关键词： cryo-EM protein–liposome complexes liposome signal subtraction 2D classification averaging

The autophagy protein Atg9 functions in glia and contributes to parkinsonian symptoms in a Drosophila model of Parkinson’s disease

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Neural Regeneration Research 2024年第5期19卷 1150-1155页

作者： Shuanglong Yi Linfang Wang Margaret S.Ho Shiping Zhang school of life Science and technology ShanghaiTech UniversityShanghaiChina College of life sciences University of Chinese Academy of SciencesBeijingChina

Parkinson’s disease is a progressive neurodegenerative disease characterized by motor deficits,dopaminergic neuron loss,and brain accumulation ofα-synuclein aggregates called Lewy *** in protein degradation pathways,such as autophagy,has been demonstrated in neurons as a critical mechanism for eliminating protein aggregates in Parkinson’s ***,it is less well understood how protein aggregates are eliminated in glia,the other cell type in the *** the present study,we show that autophagy-related gene 9(Atg9),the only transmembrane protein in the autophagy machinery,is highly expressed in Drosophila glia from adult *** from immunostaining and live cell imaging analysis reveal that a portion of Atg9 localizes to the trans-Golgi network,autophagosomes,and lysosomes in ***9 is persistently in contact with these *** glial atg9 reduces the number of omegasomes and autophagosomes,and impairs autophagic substrate *** suggests that glial Atg9 participates in the early steps of autophagy,and hence the control of autophagic ***,loss of glial atg9 induces parkinsonian symptoms in Drosophila including progressive loss of dopaminergic neurons,locomotion deficits,and glial *** findings identify a functional role of Atg9 in glial autophagy and establish a potential link between glial autophagy and Parkinson’s *** results may provide new insights on the underlying mechanism of Parkinson’s disease.

关键词： Atg9 autophagy glia Parkinson’s disease

How variable progenitor clones construct a largely invariant neocortex

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National Science Review 2024年第1期11卷 129-144页

作者： Zhongfu Shen Jiajun Yang Qiangqiang Zhang Kuiyu Wang Xiaohui Lv Xiaolin Hu Jian Ma Song-Hai Shi New Cornerstone Science Laboratory IDG/McG overn Institute for Brain Research Tsinghua-Peking Center for Life Sciences Beijing Frontier Research Center for Biological Structure School of Life Sciences Tsinghua University Department of Computer sciences Tsinghua University College of life sciences Nankai University Chinese Institute for Brain Research

The neocortex contains a vast collection of diverse neurons organized into distinct layers. While nearly all neocortical neurons are generated by radial glial progenitors(RGPs), it remains largely unclear how a complex yet organized neocortex is constructed reliably and robustly. Here, we show that the division behavior and neuronal output of RGPs are highly constrained with patterned variabilities to support the reliable and robust construction of the mouse neocortex. The neurogenic process of RGPs can be well-approximated by a consistent Poisson-like process unfolding over time, producing deep to superficial layer neurons progressively. The exact neuronal outputs regarding layer occupation are variable; yet, this variability is constrained systematically to support all layer formation, largely reflecting the variable intermediate progenitor generation and RGP neurogenic entry and exit timing differences. Together, these results define the fundamental features of neocortical neurogenesis with a balanced reliability and variability for the construction of the complex neocortex.

关键词： neocortex radial glial progenitor intermediate progenitor neurogenesis clonal variability layer formation

Visualization of cristae and mtDNA interactions via STED nanoscopy using a low saturation power probe

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Light(Science & Applications) 2024年第9期13卷 1741-1756页

作者： Wei Ren Xichuan Ge Meiqi Li Jing Sun Shiyi Li Shu Gao Chunyan Shan Baoxiang Gao Peng Xi Department of Biomedical Engineering National Biomedical Imaging CenterCollege of Future TechnologyPeking UniversityBeijing 100871China Key Laboratory of Analytical Science and technology of Hebei Province College of Chemistry and Material ScienceHebei UniversityBaoding 071002China school of life sciences Peking UniversityBeijing 100871China National center for protein sciences Peking UniversityBeijing 100871China

Mitochondria are crucial organelles closely associated with cellular metabolism and *** DNA(mtDNA)encodes a variety of transcripts and proteins essential for cellular ***,the interaction between the inner membrane(IM)and mtDNA remains elusive due to the limitations in spatiotemporal resolution offered by conventional microscopy and the absence of suitable in vivo probes specifically targeting the ***,we have developed a novel fluorescence probe called HBmito Crimson,characterized by exceptional photostability,fluorogenicity within lipid membranes,and low saturation *** successfully achieved over 500 frames of lowpower stimulated emission depletion microscopy(STED)imaging to visualize the IM dynamics,with a spatial resolution of 40 *** utilizing dual-color imaging of the IM and mtDNA,it has been uncovered that mtDNA tends to habitat at mitochondrial tips or branch points,exhibiting an overall spatially uniform ***,the dynamics of mitochondria are intricately associated with the positioning of mtDNA,and fusion consistently occurs in close proximity to mtDNA to minimize pressure during cristae *** healthy cells,>66%of the mitochondria are Class III(i.e.,mitochondria>5μm or with>12 cristae),while it dropped to<18%in *** dynamics,orchestrated by cristae remodeling,foster the even distribution of ***,in conditions of apoptosis and ferroptosis where the cristae structure is compromised,mtDNA distribution becomes *** findings,achieved with unprecedented spatiotemporal resolution,reveal the intricate interplay between cristae and mtDNA and provide insights into the driving forces behind mtDNA distribution.

关键词： metabolism saturation utilizing

Multi-region calcium imaging in freely behaving mice with ultra-compact head-mounted fluorescence microscopes

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National Science Review 2024年第1期11卷 161-173页

作者： Feng Xue Fei Li Ke-ming Zhang Lufeng Ding Yang Wang Xingtao Zhao Fang Xu Danke Zhang Mingzhai Sun Pak-Ming Lau Qingyuan Zhu Pengcheng Zhou Guo-Qiang Bi Department of Precision Machinery and Precision Instruments University of Science and Technology of China Hefei National Laboratory for Physical sciences at the Microscale University of Science and Technology of China Interdisciplinary center for Brain Information the Brain Cognition and Brain Disease Institute Shenzhen Institute of Advanced Technology Chinese Academy of Sciences Faculty of life and Health sciences Shenzhen Institutes of Advanced Technology Chinese Academy of Sciences school of life sciences University of Science and Technology of China Institute of Semiconductors Chinese Academy of Sciences Department of Modern life sciences and Biotecnology Xiongan Institute of Innovation Xiongan New Area Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions Suzhou Institute for Advanced Research University of Science and Technology of China Institute of Artiflcial Intelligence Hefei Comprehensive National Science Center

To investigate the circuit-level neural mechanisms of behavior, simultaneous imaging of neuronal activity in multiple cortical and subcortical regions is highly desired. Miniature head-mounted microscopes offer the capability of calcium imaging in freely behaving animals. However, implanting multiple microscopes on a mouse brain remains challenging due to space constraints and the cumbersome weight of the ***, we present TINIscope, a Tightly Integrated Neuronal Imaging microscope optimized for electronic and opto-mechanical design. With its compact and lightweight design of 0.43 g, TINIscope enables unprecedented simultaneous imaging of behavior-relevant activity in up to four brain regions in ***-of-concept experiments with TINIscope recorded over 1000 neurons in four hippocampal subregions and revealed concurrent activity patterns spanning across these regions. Moreover, we explored potential multi-modal experimental designs by integrating additional modules for optogenetics, electrical stimulation or local field potential recordings. Overall, TINIscope represents a timely and indispensable tool for studying the brain-wide interregional coordination that underlies unrestrained behaviors.

关键词： miniature microscope multiple sites calcium imaging multimodal integration hippocampus

Interplant communication:an emerging battlefield in plant-aphid-virus interactions

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Science China(life sciences) 2024年第5期67卷 1069-1071页

作者： Yunjing Wang Qian Gong Yule Liu MOE Key Laboratory of Bioinformatics and center for Plant Biology School of Life SciencesTsinghua UniversityBeijing 100084China Tsinghua-Peking center for life sciences Beijing 100084China

Plants constantly face threats from various organisms,including pathogens and *** the pathogens,plant viruses are obligate intracellular parasites that infect plants and cause serious crop yield losses.

关键词： plant Plants communication