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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 grazing alleviates aeolian erosion–deposition effects on microbial communities in a semi-arid grassland

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Ecological Processes 2024年第2期13卷 148-164页

作者： Mingming Cui Biqi Bao Yipu Wu Nan Hui Mai-He Li Shiqiang Wan Shijie Han Feirong Ren Junqiang Zheng International joint Research Laboratory for Global Change Ecology School of Life Sciences Henan University Yellow River Floodplain Ecosystems Research Station School of Life Sciences Henan University school of Agriculture and Biology Shanghai Jiao Tong University Forest Dynam-ics Swiss Federal Institute for Forest Snow and Landscape Research WSL Key Laboratory of Geographical Processes and Ecological Security in Changbai Mountains Ministry of Education School of Geographical Sciences Northeast Normal University school of life Science Hebei University

Background Soil erosion affects the stability of terrestrial ecosystems and ecosystem services by directly or indirectly impacting the cycling of soil materials and energy and reducing the fertility of grassland soils. However, research on microbial adaptation to grazing and soil erosion is limited, particularly in relation to grassland ecosystem restora-tion. Here, we assess microbial communities subjected to simulated soil erosion and grazing in a semi-arid grassland of Inner Mongolia, *** No significant change was observed in soil variables. However, the structure of the soil microbial commu-nity underwent significant changes as a result of soil erosion and soil erosion plus grazing, leading to a significant increase in the relative abundance of Cyanobacteria(116.80% vs 116.38%). Wind erosion and deposition contributed to an increase in the network complexity of soil bacterial and fungal communities. However, much of this effect was alleviated by grazing. Simultaneously, aeolian processes and grazing regulate soil microbial community assembly, leading to inconsistent patterns of change in bacterial and fungal communities. Under wind erosion and deposi-tion, the relative contribution of deterministic processes(4.44% vs 31.11%) in bacterial communities increased, while the relative contribution of stochastic processes(2.23% vs 20%) in fungal communities reduced. Grazing resulted in a decrease in the relative contribution of deterministic processes(8.89%) in the bacterial community and an increase in the relative contribution of stochastic processes(8.89%) in the fungal *** This study presents a comprehensive investigation of the response of soil microbial communities to aeolian erosion–deposition and grazing in a semi-arid grassland. Our findings indicate that microbial communities in the semi-arid grassland show resistance to external disturbances and that light grazing mitigates the effects of aeo-lian erosion–deposition on microbial co

关键词： Semi-arid grassland Erosion Grazing Soil microbial community network Deposition

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

Physiological and pathological functions of circular RNAs in the nervous system

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Neural Regeneration Research 2024年第2期19卷 342-349页

作者： Min Zhou Shi Li Chuan Huang school of life sciences Chongqing UniversityChongqingChina

Circular RNAs(circRNAs)are a class of covalently closed single-stranded RNAs that are expressed during the development of specific cells and *** play crucial roles in physiological and pathological processes by sponging microRNAs,modulating gene transcription,controlling the activity of certain RNA-binding proteins,and producing functional peptides.A key focus of research at present is the functionality of circRNAs in the nervous system and several advances have emerged over the last 2 ***,the precise role of circRNAs in the nervous system has yet to be comprehensively *** this review,we first summarize the recently described roles of circRNAs in brain development,maturity,and ***,we focus on the involvement of circRNAs in various diseases of the central nervous system,such as brain cancer,chronic neurodegenerative diseases,acute injuries of the nervous system,and neuropathic pain.A better understanding of the functionality of circRNAs will help us to develop potential diagnostic,prognostic,and therapeutic strategies to treat diseases of the nervous system.

关键词： Alzheimer’s disease amyotrophic lateral sclerosis brain development circRNAs neuropathic pain Parkinson’s disease

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

Inhibition mechanisms of CRISPR-Cas9 by AcrⅡA25.1 and AcrⅡA32

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Science China(life sciences) 2024年第9期67卷 1781-1791页

作者： Jianlin Zheng Yuwei Zhu Tengjin Huang Wenbo Gao Jiale He Zhiwei Huang HIT Center for life sciences School of Life Science and TechnologyHarbin Institute of TechnologyHarbin 150080China Westlake Center for Genome Editing Westlake Laboratory of Life Sciences and BiomedicineSchool of Life SciencesWestlake UniversityHangzhou 310024China

In the ongoing arms race between bacteria and bacteriophages,bacteriophages have evolved anti-CRISPR proteins to counteract bacterial CRISPR-Cas ***,AcrⅡA25.1 and AcrⅡA32 have been found to effectively inhibit the activity of Spy Cas9 both in bacterial and human ***,their molecular mechanisms remain ***,we report the cryo-electron microscopy structures of ternary complexes formed by AcrⅡA25.1 and AcrⅡA32 bound to Spy Cas9-sg *** structural analysis and biochemical experiments,we revealed that AcrⅡA25.1 and AcrⅡA32 recognize a novel,previously-unidentified anti-CRISPR binding site on Spy *** found that both AcrⅡA25.1 and AcrⅡA32 directly interact with the WED domain,where they spatially obstruct conformational changes of the WED and PI domains,thereby inhibiting Spy Cas9 from recognizing protospacer adjacent motif(PAM)and unwinding double-stranded *** addition,they may inhibit nuclease activity by blocking the dynamic conformational changes of the Spy Cas9 surveillance *** summary,our data elucidate the inhibition mechanisms of two new anti-CRISPR proteins,provide new strategies for the modulation of Spy Cas9 activity,and expand our understanding of the diversity of anti-CRISPR protein inhibition mechanisms.

关键词： anti-CRISPR cryo-EM Spy Cas9 AcrⅡA25.1 AcrⅡA32 inhibiton

Impact of tourism on the vigilance behavior of the Cyprus rock agama(Laudakia cypriaca)

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Integrative Zoology 2024年第3期19卷 577-583页

作者： Artur GOLAWSKI Iris CHARALAMBIDOU Sylwia GOLAWSKA Faculty of sciences Siedlce University of Natural Sciences and HumanitiesSiedlcePolandand Department of life sciences School of Life and Health SciencesUniversity of NicosiaNicosiaCyprus

Escape behavior is a common antipredator strategy among wild ***,we investigated the effect of four factors on the vigilance behavior of the endemic Cyprus rock agama(Laudakia cypriaca).Flight initiation distance(FID,the minimum distance to which an observer can approach a lizard before it flees)was measured in relation to the type of location(tourist *** area),the observer’s starting distance,air temperature,and substrate *** collected data for 39 agamas in tourist areas and 34 of these lizards in nontourist *** a whole,the mean starting distance was 10.5 m and the FID was 3.6 *** average substrate temperature was 34.0℃ and the average air temperature 29.6℃.Only the type of area affected the agamas’escape decisions with FID being 1.8 m shorter in tourist areas than in nontourist areas(2.7 m vs.4.5 m).This is probably due to the habituation of lizards to the presence of humans in the former *** study shows that tourism strongly affects the behavior of lizards,which may have consequences for the functioning of the *** can increase the safety of lizards by creating a human shield to deter *** the tourist season is over,lizards may become more vulnerable to predators.

关键词： anthropogenic disturbance flight initiation distance lizards tourism vigilance behavior

PRRSV degrades MDA5 via dual autophagy receptors P62 and CCT2 to evade antiviral innate immunity

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Virologica Sinica 2024年第2期39卷 264-276页

作者： Ruiqi Sun Yanyu Guo Lilin Zhang Huixia Zhang Boxuan Yin Xiaoyang Li Changyan Li Liu Yang Lei Zhang Zexing Li Jinhai Huang school of life sciences Tianjin UniversityTianjin300072China

Porcine reproductive and respiratory syndrome virus(PRRSV)is a major economically devastating pathogen that has evolved various strategies to evade innate *** of antiviral interferon largely promotes PRRSV immunoevasion by utilizing cytoplasmic melanoma differentiation-associated gene 5(MDA5),a receptor that senses viral *** this study,the downregulated transcription and expression levels of porcine MDA5 in PRRSV infection were observed,and the detailed mechanisms were *** found that the interaction between P62 and MDA5 is enhanced due to two factors:the phosphorylation modification of the autophagic receptor P62 by the upregulated kinase CK2αand the K63 ubiquitination of porcine MDA5 catalyzed by the E3 ubiquitinase TRIM21 in PRRSV-infected *** a result of these modifications,the classic P62-mediated autophagy is ***,porcine MDA5 interacts with the chaperonin containing TCP1 subunit 2(CCT2),which is enhanced by PRRSV *** interaction promotes the aggregate formation and autophagic clearance of MDA5-CCT2-nsp3 independently of *** summary,enhanced MDA5 degradation occurs in PRRSV infection via two autophagic pathways:the binding of MDA5 with the autophagy receptor P62 and the aggrephagy receptor CCT2,leading to intense innate immune *** research reveals a novel mechanism of immune evasion in PRRSV infection and provides fundamental insights for the development of new vaccines or therapeutic strategies.

关键词： Porcine reproductive and respiratory syndrome virus(PRRSV) nsp3 Autophagy MDA5

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