Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an effic...Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.展开更多
In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men a...In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men and two women) and four healthy male rats was imaged during diastolic and systolic phases on the mid-ventricle level. The RS and radial SR of the left ventricle were calculated at the mid-ventricular level of the cardiac cycle. The peak RS for rat and human heart was found to be 46.8 ± 0.68 and 40.7 ± 1.44, respectively, and it occurred at 40% of the cardiac cycle for both human and rat hearts. The peak systolic and diastolic radial SR for human heart was 1.10 ± 0.08 s- 1 and - 1.78 ± 0.02 s- 1, respectively, while it was 4.25 ± 0.02 s- 1 and - 5.16 ± 0.23 s- 1, respectively for rat heart. The results show that tMRI data can be used to characterize the cardiac function during systolic and diastolic phases of the cardiac cycle, and as a result, it can be used to evaluate the cardiac motion by calculating its RS and radial SR at different locations of the cardiac wall during both diastolic and systolic phases. This study also approves the validity of the tagged MRI data to accurately describe the radial cardiac motion.展开更多
Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which gene...Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which genetic transformation is not routine. Two Ds transposon flanking sequences(TNPs), TNP-29(27.4 c M(centi Morgan)) and TNP-79(70.3 c M), were mapped in the vicinity of a malting quality QTL located on chromosome 4H of barley. Reactivation of the Ds transposon sequence from these TNP lines led to the identification of genes in the malting QTL regions. Several Ds(dissociation) lines were generated by crossing TNP-29 and TNP-79 with an Ac TPase(activator) expressing line(25-B), and F2 progenies were subsequently screened for Ds insertions at new locations. To further characterize these Ds mutants, we mapped the new Ds flanking sequences on a barley genetic map and found that 29% of Ds were located in regions associated with the malting QTL located on chromosome 4H and in close proximity to other important malting-associated QTL across the barley chromosome. Using a sequence based approach, a linkage map was generated that confirmed the position of Ds loci in the barley genome map. Locating these Ds loci on the barley map opens avenues to dissect important malting QTL for facilitating identification of candidate malting genes.展开更多
Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunopreci...Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.展开更多
Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal ...Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.展开更多
The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions an...The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions and protons in the final state were detected with the Neutral Kaon Spectrometer （NKS2）. Photon energies were obtained from the reaction of d（γ,π-pp）. The derived photon energy was consistent with the design of the tagger system and the previous measurement using electron-positron pair production. The consistency demonstrates the performance of NKS2 and the capability of the photon energy calibration using d（γ,π-pp）.展开更多
Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numb...Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numbers and the central limit theorem for the tagged particle. The proof of the central limit theorem is based on the method of martingale decomposition with a sector condition.展开更多
We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its...We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its position in Zd,denoted by Xt. Note that the whole space is not homogeneous. We define the environment process viewed from the tagged particle, of which Xt can be expressed as a functional. It is called the tagged particle process. We show the ergodicity of the tagged particle process, then prove the strong law of large numbers. Furthermore, we show the central limit theorem of Xt provided the zero-mean condition.展开更多
Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,ca...Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,cannot be incorporated with GFP tag as a large fragment.It was recently reported that protein genetically inserted with a smaller size tetracysteine(TC)tag could be specially labeled by a biarsenicalfluorescent dye in living cells.In this study,we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion.TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis.Western blot and enzyme-linked immu-nosorbent assay(ELISA)analysis showed that the TC-tagged core protein,hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)could be expressed in cells transfected with the recombinant HBV vector,which is similar to the cells transfected with wild-type HBV vector.Reverse transcription-polymerase chain reaction(RT-PCR)and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree,but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins.Taken together,the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to befluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.展开更多
The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-s...The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411(HA-tagged Ssp411)mouse model.This model is instrumental in probing the expression and function of Ssp411.Our research revealed that Ssp411 is expressed in the round spermatids,elongating spermatids,elongated spermatids,and epididymal spermatozoa.The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis.Nevertheless,rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions.Ssp411 is not detectable in metaphase Ⅱ(MⅡ)oocytes,zygotes,or 2-cell stage embryos,highlighting its intricate role in early embryonic development.These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP,fostering groundbreaking advancements in the f ields of spermiogenesis and reproductive biology.展开更多
Utilizing small molecules as markers for specific cells or organs within biosystems is a crucial approach for studying and regulating physiological processes. However, current tagging strategies, due to the presence o...Utilizing small molecules as markers for specific cells or organs within biosystems is a crucial approach for studying and regulating physiological processes. However, current tagging strategies, due to the presence of exposed highly reactive groups, suffer from drawbacks such as low tagging efficiency or insufficient spatial specificity, thereby diminishing their expected effectiveness. Consequently, there is a pressing need to develop a strategy capable of in situ labeling of active groups in response to cellular or in vivo stimuli, ensuring both high tagging efficiency and spatial specificity. In this work, we devised a strategy for releasing aldehyde groups activated by hypochlorous acid(HOCl). Compounds synthesized through this strategy can release the fiuorophore methylene blue(MB) and aldehyde-based compounds upon HOCl activation. Given high reactivity of the released aldehyde group, it can effectively interact with macromolecules in biological systems, facilitating tagging and enabling prolonged imaging. To validate this concept, we further incorporated a naphthalimide structure with stable light emission to create SW-110. SW-110 can specifically respond to in vitro and endogenous HOCl, when release MB, it also releases naphthalimide fiuorophore with highly reactive aldehyde group for tagging within cells. This strategy provides a simple but efficient strategy for proximity tagging in situ.展开更多
基金the Animal Ethics Committee of the Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences(SYXK-2020-0010).
文摘Influenza A viruses(IAVs)possess variable pathogenic potency causing great economic losses in the poultry industry worldwide and threatening public health.The control of IAV epidemics desperately necessitates an efficient platform for screening antiviral compounds and evaluating vaccine efficacy.In this study,we utilized the H9N2 subtype IAV as the working model.An 11-amino-acid HiBiT tag,derived from NanoLuc luciferase,was incorporated into the flexible linker region of the NS1 protein.Subsequently,the recombinant HiBiT-tagged virus was rescued.The recombinant virus exhibited high genetic stability and similar virological characteristics to the parental virus,both in vitro and in vivo.Particularly importantly,the replication profile of the HiBiT-tagged virus can be easily measured using the Nano-Glo assay system,achieving an efficient screening platform.Based on this platform,we have developed assays with both convenience and efficiency for screening antiviral reagents,evaluating immunization efficacy,and measuring neutralizing antibodies.
文摘In this paper, the Radial Strain (RS) and Strain Rate (SR) was calculated using tagged MRI (tMRI) data. Using tagged magnetic resonance imaging (tMRI), the left ventricle short axis of five healthy adults (three men and two women) and four healthy male rats was imaged during diastolic and systolic phases on the mid-ventricle level. The RS and radial SR of the left ventricle were calculated at the mid-ventricular level of the cardiac cycle. The peak RS for rat and human heart was found to be 46.8 ± 0.68 and 40.7 ± 1.44, respectively, and it occurred at 40% of the cardiac cycle for both human and rat hearts. The peak systolic and diastolic radial SR for human heart was 1.10 ± 0.08 s- 1 and - 1.78 ± 0.02 s- 1, respectively, while it was 4.25 ± 0.02 s- 1 and - 5.16 ± 0.23 s- 1, respectively for rat heart. The results show that tMRI data can be used to characterize the cardiac function during systolic and diastolic phases of the cardiac cycle, and as a result, it can be used to evaluate the cardiac motion by calculating its RS and radial SR at different locations of the cardiac wall during both diastolic and systolic phases. This study also approves the validity of the tagged MRI data to accurately describe the radial cardiac motion.
基金Funding for this project was provided by Barley Malting and Brewing Research Institute (grant number: 217248)
文摘Among various functional genomics tools used to characterize genes in plants, transposonbased mutagenesis approaches offer great potential, especially in barley and wheat, which possess large genomes and in which genetic transformation is not routine. Two Ds transposon flanking sequences(TNPs), TNP-29(27.4 c M(centi Morgan)) and TNP-79(70.3 c M), were mapped in the vicinity of a malting quality QTL located on chromosome 4H of barley. Reactivation of the Ds transposon sequence from these TNP lines led to the identification of genes in the malting QTL regions. Several Ds(dissociation) lines were generated by crossing TNP-29 and TNP-79 with an Ac TPase(activator) expressing line(25-B), and F2 progenies were subsequently screened for Ds insertions at new locations. To further characterize these Ds mutants, we mapped the new Ds flanking sequences on a barley genetic map and found that 29% of Ds were located in regions associated with the malting QTL located on chromosome 4H and in close proximity to other important malting-associated QTL across the barley chromosome. Using a sequence based approach, a linkage map was generated that confirmed the position of Ds loci in the barley genome map. Locating these Ds loci on the barley map opens avenues to dissect important malting QTL for facilitating identification of candidate malting genes.
文摘Comprehensive understanding of mechanisms of epigenetic regulation requires identification of molecules bound to genomic regions of interest in vivo. We have developed a novel method, insertional chromatin immunoprecipitatin (iChIP), to isolate specific genomic regions retaining molecular interaction in order to perform non-biased identification of interacting molecules in vivo. Here, we developed a second-generation tagged LexA DNA-binding domain, 3xFNLDD, for the iChIP analysis. 3xFNLDD consists of 3 x FLAG tags, a nuclear localization signal (NLS), the DNA-binding domain (DB) and the dimerization domain of the LexA protein. Expression of 3xFNLDD can be detected by immunoblot analysis as well as flowcytometry. We showed that iChIP using 3xFNLDD is able to consistently isolate more than 10% of input genomic DNA, several-fold more efficient compared to the first-generation tagged LexA DB. 3xFNLDD would be a useful tool to perform the iChIP analysis for locus-specific biochemical epigenetics.
基金supported by National Natural Science Foundation of China(T2122002,82361148715,22077079,82204104)National Key R&D Program of China(2022YFC2601700,2022YFF0710202 and 2022YFA1104200)+4 种基金Princess Nourah bint Abdulrahman University Researchers Supporting Project number(PNURSP2024R122)Shanghai Municipal Science and Technology Projects(22Z510202478)Shanghai Municipal Education Commission Projects(21SG10,ZXWH1082101)Shanghai Jiao Tong University Projects(YG2021ZD19)Shanghai University of Medicine&Health Sciences Project(AMSCP-24-07-01).
文摘Mass cytometry(cytometry by time-of-flight(CyTOF))and imaging mass cytometry(IMC)are transformative technologies that combine flow cytometry principles with time-of-flight mass spectrometry(TOF-MS).By employing metal isotope-tagged antibodies instead of fluorophores,these techniques overcome spectral overlap limitations and enable high-dimensional,compensation-free analysis of complex biological systems at single-cell resolution.The performance of CyTOF and IMC critically depends on advanced nanomaterials labeled with stable metal isotopes,which are essential for improving sensitivity and multiplexing capacity.This review systematically discusses the design principles,synthesis methods,and functionalization strategies of mass-tagged nanomaterials tailored for CyTOF(e.g.,cell suspension analysis)and IMC(e.g.,spatial proteomics of tissue sections).We highlight their impactful applications in biomedicine,including proteomics,immunology,oncology,and neuroscience,emphasizing their roles in disease diagnosis,targeted drug development,and singlecell analysis.Despite these advancements,challenges such as nanomaterial biocompatibility,clinical scalability,and artificial intelligence(AI)-driven design are discussed,providing a roadmap for future research in personalized medicine and theranostics.
基金Supported by a Grant-in-Aid (16GS0201) for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan
文摘The energy of tagged photons, which were provided from the internal photon tagging system of the Laboratory of Nuclear Science, Tohoku University, has been calibrated using the d（γ,π-pp） reaction. Charged pions and protons in the final state were detected with the Neutral Kaon Spectrometer （NKS2）. Photon energies were obtained from the reaction of d（γ,π-pp）. The derived photon energy was consistent with the design of the tagger system and the previous measurement using electron-positron pair production. The consistency demonstrates the performance of NKS2 and the capability of the photon energy calibration using d（γ,π-pp）.
文摘Stirring-exclusion processes are exclusion processes with particles being stirred. We investigate a tagged particle among a Bernoulli product environment measure on the lattice Zd. We show the strong law of large numbers and the central limit theorem for the tagged particle. The proof of the central limit theorem is based on the method of martingale decomposition with a sector condition.
基金supported by National Natural Science Foundation of China(Grant No.11371040)
文摘We investigate a tagged particle in the exclusion processes on {1,..., N }×Zd, with different densities in different levels {k} × Zd, ? k. Ignoring the level the tagged particle lying in, we only concern its position in Zd,denoted by Xt. Note that the whole space is not homogeneous. We define the environment process viewed from the tagged particle, of which Xt can be expressed as a functional. It is called the tagged particle process. We show the ergodicity of the tagged particle process, then prove the strong law of large numbers. Furthermore, we show the central limit theorem of Xt provided the zero-mean condition.
基金supported in part by the National Natural Science Foundation of China(Grant Nos.30872237 and 30600277)National Key Basic Research Program of China(973 Program,No.2007CB512900)Doctoral Fund of Ministry of Education of China(No.20070487007).
文摘Virus tagged with greenfluorescent protein(GFP)contributes to the visualization and study of the virus in living cells.However,the hepatitis B virus(HBV)particle,which is a compact virion with limited internal space,cannot be incorporated with GFP tag as a large fragment.It was recently reported that protein genetically inserted with a smaller size tetracysteine(TC)tag could be specially labeled by a biarsenicalfluorescent dye in living cells.In this study,we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion.TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis.Western blot and enzyme-linked immu-nosorbent assay(ELISA)analysis showed that the TC-tagged core protein,hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)could be expressed in cells transfected with the recombinant HBV vector,which is similar to the cells transfected with wild-type HBV vector.Reverse transcription-polymerase chain reaction(RT-PCR)and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree,but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins.Taken together,the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to befluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.
基金the support from the National Natural Science Foundation of China(No.32070849)The Foundation of Science and Technology Commission of Shanghai Municipality(No.22DX1900400)+1 种基金Science and Technology Commission of Shanghai Municipality(No.23JC1403803)Shanghai Municipal Science and Technology Commission Targeted Funding Project(No.22DX1900400).
文摘The genome tagging project(GTP)plays a pivotal role in addressing a critical gap in the understanding of protein functions.Within this framework,we successfully generated a human influenza hemagglutinin-tagged sperm-specific protein 411(HA-tagged Ssp411)mouse model.This model is instrumental in probing the expression and function of Ssp411.Our research revealed that Ssp411 is expressed in the round spermatids,elongating spermatids,elongated spermatids,and epididymal spermatozoa.The comprehensive examination of the distribution of Ssp411 in these germ cells offers new perspectives on its involvement in spermiogenesis.Nevertheless,rigorous further inquiry is imperative to elucidate the precise mechanistic underpinnings of these functions.Ssp411 is not detectable in metaphase Ⅱ(MⅡ)oocytes,zygotes,or 2-cell stage embryos,highlighting its intricate role in early embryonic development.These findings not only advance our understanding of the role of Ssp411 in reproductive physiology but also significantly contribute to the overarching goals of the GTP,fostering groundbreaking advancements in the f ields of spermiogenesis and reproductive biology.
基金financially supported by the National Natural Science Foundation of China (Nos. 22177019, 22377010, 22371038)State Key Laboratory for Modification of Chemical Fibers and Polymer Materials (No. KF2206)。
文摘Utilizing small molecules as markers for specific cells or organs within biosystems is a crucial approach for studying and regulating physiological processes. However, current tagging strategies, due to the presence of exposed highly reactive groups, suffer from drawbacks such as low tagging efficiency or insufficient spatial specificity, thereby diminishing their expected effectiveness. Consequently, there is a pressing need to develop a strategy capable of in situ labeling of active groups in response to cellular or in vivo stimuli, ensuring both high tagging efficiency and spatial specificity. In this work, we devised a strategy for releasing aldehyde groups activated by hypochlorous acid(HOCl). Compounds synthesized through this strategy can release the fiuorophore methylene blue(MB) and aldehyde-based compounds upon HOCl activation. Given high reactivity of the released aldehyde group, it can effectively interact with macromolecules in biological systems, facilitating tagging and enabling prolonged imaging. To validate this concept, we further incorporated a naphthalimide structure with stable light emission to create SW-110. SW-110 can specifically respond to in vitro and endogenous HOCl, when release MB, it also releases naphthalimide fiuorophore with highly reactive aldehyde group for tagging within cells. This strategy provides a simple but efficient strategy for proximity tagging in situ.
