Based on sequencing of part clones in a root subtractive cDNA library, an expressed sequence tag （EST） sharing high similarity to a rice C2H2 zinc finger transcription factor （ZFP15） was obtained in wheat. Through...Based on sequencing of part clones in a root subtractive cDNA library, an expressed sequence tag （EST） sharing high similarity to a rice C2H2 zinc finger transcription factor （ZFP15） was obtained in wheat. Through bioinformatics approach, the wheat C2H2-type ZFP gene referred to TaZFP15 has been identified and characterized. As a full-length cDNA of 670 bp, TaZFP15 has an open reading frame of 408 bp and encodes a 135-aa polypeptide. TaZFP15 contains two C2H2 zinc finger domains and each one has a conserved motif QALGGH. The typical L-box, generally identified in the C2H2 type transcription factors, has also been found in TaZFP15. Phylogenetic analysis suggested that TaZFP15 shares high similarities with rice ZFP15 （GenBank accession no. AY286473）, maize ZFP （GenBank accession no. NM_001159094） and a subset of other zinc-finger transcription factor genes in plant species. The expression of TaZFP15 was up-regulated by starved-Pi stress, showing a pattern to be gradually elevated along with the progression of the Pi-stress in a 23-h treatment regime. Similarly, the transcripts of TaZFP15 in roots were also induced by nitrogen deficiency, and abiotic stresses of drought and salinity. No responses of TaZFP15 were detected in roots to nutrition deficiencies of P, Zn, and Ca, and the external treatment of abscisic acid （ABA）. TaZFP15 could be specifically amplified in genome A, B, and D, and without variability in the sequences, suggesting that TaZFP15 has multi-copies in the homologous hexaploid species. Transgenic analysis in tobacco revealed that up-regulation of TaZFP15 could significantly improve plant dry mass accumulation via increasing the plant phosphorus acquisition capacity under Pi-deficiency condition. The results suggested that TaZFP15 is involved in mediation of signal transductions of diverse external stresses.展开更多
The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P...The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P. euphratica. Northern hybridization revealed that its expression was induced under drought and salt stress conditions. To examine its function, cDNA of the PSTZ gene, driven by the cauliflower mosaic virus 35S promoter, was cloned into a plant expression vector pBin438 and introduced into tobacco plants. Transgenic tobacco showed an enhanced salt tolerance, suggesting that PSTZ may play a role in plant responsiveness to salt stress.展开更多
Ikaros represents a zinc-finger protein family important for lymphocyte development and certain other physiological processes. The number of family members is large, with alternative splicing producing various additio...Ikaros represents a zinc-finger protein family important for lymphocyte development and certain other physiological processes. The number of family members is large, with alternative splicing producing various additional isoforms from each of the five homologous genes in the family. The functional forms of Ikaros proteins could be even more diverse due to protein–protein interactions readily established between family members. Emerging evidence suggests that targeting Ikaros proteins is feasible and effective in therapeutic applications, although the exact roles of Ikaros proteins remain elusive within the intricate regulatory networks in which they are involved. In this review we collect existing knowledge as to the functions, regulatory pathways, and molecular mechanisms of this family of proteins in an attempt to gain a better understanding through the comparison of activities and interactions among family members.展开更多
An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. c...An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.展开更多
CCCTC-binding factor(CTCF) is a zinc-finger protein, serving an important part in the genome architecture as well as some biochemical processes. Over 70,000 CTCF binding DNA sites have been detected genome-wide, and m...CCCTC-binding factor(CTCF) is a zinc-finger protein, serving an important part in the genome architecture as well as some biochemical processes. Over 70,000 CTCF binding DNA sites have been detected genome-wide, and most anchors of chromatin loops are demarcated with the CTCF binding.Various protein or RNA molecules interact with DNA-bound CTCF to conduct different biological functions, and potentially the interfaces between CTCF and its cofactors can be targets for drug development. Here we identify the effective region of CTCF in DNA recognition, which defines the exposed CTCF surface feature for the interaction of cofactors. While the zinc-finger region contributes the most in DNA association, its binding affinity varies based on different DNA sequences. To investigate the effectiveness of individual zinc-fingers, the key residues are mutated to inactivate the DNA binding ability, while the finger configuration and the spacing between fingers are preserved. The strategy is proved to be successful, while clear differences are observed in the DNA binding affinities among the 11 finger mutants and the result is consistent to previous studies in general. With the help of inactivated finger mutants, we identify the ineffective fingers and the dominant effective fingers, which form distinctive patterns on different DNA targets.展开更多
RING is a really interesting new gene which plays important regulatory roles in many developmental processes as well as in plant-environment interactions. In the present report, the Zm RHCP1 gene encoding a putative R...RING is a really interesting new gene which plays important regulatory roles in many developmental processes as well as in plant-environment interactions. In the present report, the Zm RHCP1 gene encoding a putative RING-HC protein was isolated from maize and characterized. The Zm RHCP1 protein contained 310 amino acid residues with a conserved RINGHC zinc-finger motif and two transmembrane（TM） domains. Zm RHCP1 was expressed ubiquitously in various organs（root, stem, leaf, seedling, immature ear, and tassel）, but its transcript levels were higher in vegetative organs than in reproductive organs. Moreover, the expression pattern of Zm RHCP1 in brace roots indicated that Zm RHCP1 functions in brace root initiation. In addition, Zm RHCP1 expression was regulated by abiotic stresses. The expression results suggested that Zm RHCP1 plays important roles in brace root development and abiotic stress responses. The findings of the present study provide important information to help us understand the function of Zm RHCP1 in maize.展开更多
The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degra...The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degradation.ZAP directly binds to the target viral mRNA and recruits the cellular RNA degradation machinery to degrade the RNA.No significant sequence similarity or obvious common motifs have been found in the so far identified target viral mRNAs.The minimum length of the target sequence is about 500 nt long.Short workable ZAP-binding RNAs should facilitate further studies on the ZAP-RNA interaction and characterization of such RNAs may provide some insights into the underlying mechanism.In this study,we used the SELEX method to isolate ZAP-binding RNA aptamers.After 21 rounds of selection,ZAP-binding aptamers were isolated.Sequence analysis revealed that they are G-rich RNAs with predicted stem-loop structures containing conserved“GGGUGG”and“GAGGG”motifs in the loop region.Insertion of the aptamer sequence into a luciferase reporter failed to render the reporter sensitive to ZAP.However,overexpression of the aptamers modestly but significantly reduced ZAP’s antiviral activity.Substitution of the conserved motifs of the aptamers significantly impaired their ZAP-binding ability and ZAP-antagonizing activity,suggesting that the RNA sequence is important for specific interaction between ZAP and the target RNA.The aptamers identified in this report should provide useful tools to further investigate the details of the interaction between ZAP and the target RNAs.展开更多
Well-established targeted technologies to engi- neer genomes such as zinc-finger nuclease-based editing （ZFN）, transcription activator-like effector nuclease-based editing （TALEN）, and clustered regularly interspa...Well-established targeted technologies to engi- neer genomes such as zinc-finger nuclease-based editing （ZFN）, transcription activator-like effector nuclease-based editing （TALEN）, and clustered regularly interspaced short palindromic repeats and associated protein system-based editing （CRISPR/Cas） are proving to advance basic and applied research in numerous plant species. Compared with systems using ZFNs and TALENs, the most recently developed CRISPR/Cas system is more efficient due to its use of an RNA-guided nuclease to generate double-strand DNA breaks. To accelerate the applications of these technologies, we provide here a detailed overview of these systems, highlight the strengths and weaknesses of each, summarize research advances made with these technologies in model and crop plants, and discuss their applications in plant functional genomics. Such targeted approaches for genetically modifying plants will benefit agricultural production in the future.展开更多
Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platformscontributing to redefine the boundaries of modern biological research. They are composed of a non-specificcleav...Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platformscontributing to redefine the boundaries of modern biological research. They are composed of a non-specificcleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications byinducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases havebeen employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively.This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies,biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbitsusing transcription activator-like effector nucleases, and a perspective of the field.展开更多
Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1...Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1 on EC progression via the microRNA(miR)-374a-3p/zinc-finger protein(ZFX)axis.Methods:EC cell lines were collected and DLX6-AS1,miR-374a-3p,and ZFX levels in EC cell lines were detected.The EC cells were transfected with DLX6-AS1,miR-374a-3p,and ZFX constructs to examine the biological functions of EC cells.The xenograft model was established for detecting tumor growth.Rescue experiments were conducted to verify the interaction of DLX6-AS1,miR-374a-3p,and ZFX in EC cells.Results:DLX6-AS1 and ZFX levels were elevated,while miR-374a-3p exhibited a reduced level in EC cells.Silencing DLX6-AS1 and elevated miR-374a-3p expressions repressed the biological activities of EC cells.Reduced DLX6-AS1 repressed tumor development.MiR-374a-3p silencing reversed the impacts of DLX6-AS1 silencing,while ZFX overexpression abrogated the impacts of miR-374a-3p elevation on EC cell growth.Mechanically,DLX6-AS1 was found to bind to miR-374a-3p,and miR-374a-3p targeted ZFX.Conclusion:DLX6-AS1 depletion restricts the malignant phenotype of EC cells.The study might provide novel therapeutic biomarkers for EC treatment.展开更多
The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in...The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.展开更多
The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly bi...The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly binds to the viral mRNAs and recruits the RNA exosome to degrade the target RNA.In this article,we provide evidence that a DEXH box RNA helicase,DHX30,is required for optimal antiviral activity of ZAP.Pull-down and co-immunoprecipitation assays demonstrated that DHX30 and ZAP interacted with each other via their N terminal domains.Downregulation of DHX30 with shRNAs reduced ZAP’s antiviral activity.These data implicate that DHX30 is a cellular factor involved in the antiviral function of ZAP.展开更多
Inflorescence structure of rice,including the number and length of branches,and the density of the spikelet,can greatly affect the number of grains per panicle,which is one of the key factors in yield compositions.Her...Inflorescence structure of rice,including the number and length of branches,and the density of the spikelet,can greatly affect the number of grains per panicle,which is one of the key factors in yield compositions.Here we identified five allelic mutants sb1-1/2/3/4/5 that related to branch development of rice.In these mutants,the branch meristem fate was prolonged sharply,resulting in delay of transition from branches to spikelets,and then increased the numbers of branches and spikelets per panicle.SB1 encodes a nuclear RING-like domain protein of SHI/LRP/SRS family and strongly expressed in branch meristems.The results of protein interaction and chromatin immunoprecipitation further suggested that SB1 directly repressed the expression of DEP1,TAW1,MOC1 and IPA1 by interacting with a co-repressor complex to affect acetylation level of histone H3 on target regions.Thus,we proposed that SB1 is a transcription repressor of branch meristem activity by widely and negatively regulating a series of genes that maintain branch meristem fate.展开更多
Functional biological research has benefited tremendously by analyses of the phenotypes of mutant organisms which can be generated through targeted mutation of genes.In Drosophila,compared with random mutagenesis meth...Functional biological research has benefited tremendously by analyses of the phenotypes of mutant organisms which can be generated through targeted mutation of genes.In Drosophila,compared with random mutagenesis methods gene targeting has gained its popularity because it can introduce any desired mutation into a gene of interest.However,applications of gene targeting have been limited because the targeting efficiency varies with different genes,and the time and labor of targeting procedure are intensive.Nevertheless,improvement of gene targeting and development of its variant technologies have received much attention of scientists.Here we review recent progress that has been made in expanding the applications of gene targeting,which include the ϕC31 integration system and zinc-finger nucleases induced gene targeting,and new strategies that generate more efficient and reliable gene targeting.展开更多
Despite relative effectiveness of current hepatitis B therapies,there is still no curative agents available.The new emerging approaches hold promise to achieve cure and loss of hepatitis B surface antigen.Studies or c...Despite relative effectiveness of current hepatitis B therapies,there is still no curative agents available.The new emerging approaches hold promise to achieve cure and loss of hepatitis B surface antigen.Studies or clinical trials investigating new therapies remain small and either focus on patients with low viral load and without hepatotoxic injury or patients with hepatitis D co-infection,which makes it challenging to assess their effectiveness and side effect profile in hepatitis B population.展开更多
Sp1/Krüppel样因子(Sp1-like and Krüppel-like factors,Sp1/KLFs)是一组与真核细胞转录调控密切相关的锌指蛋白。Sp1/KLFs的羧基末端高度保守,含有3个串联的Cys2His2锌指结构,用于结合DNA;其氨基末端在不同的家族成员间存在...Sp1/Krüppel样因子(Sp1-like and Krüppel-like factors,Sp1/KLFs)是一组与真核细胞转录调控密切相关的锌指蛋白。Sp1/KLFs的羧基末端高度保守,含有3个串联的Cys2His2锌指结构,用于结合DNA;其氨基末端在不同的家族成员间存在较大差异,主要是通过结合辅助因子发挥转录调控作用。Sp1/KLFs的表达具有组织、细胞分布以及发育时期的特异性,它们通过调控多种富含GC或CACCC的启动子的基因的表达,参与细胞增殖、分化、凋亡和肿瘤发生、发展等多种生理、病理过程。文章综述了Sp1/KLFs的结构特征、作用机制及生物学功能。展开更多
基金supported by the National Natural Science Foundation of China (30971773)the Natural Science Foundation of Hebei Province,China (C2011204031)the Key Laboratory of Crop Growth Regulation of Hebei Province,China
文摘Based on sequencing of part clones in a root subtractive cDNA library, an expressed sequence tag （EST） sharing high similarity to a rice C2H2 zinc finger transcription factor （ZFP15） was obtained in wheat. Through bioinformatics approach, the wheat C2H2-type ZFP gene referred to TaZFP15 has been identified and characterized. As a full-length cDNA of 670 bp, TaZFP15 has an open reading frame of 408 bp and encodes a 135-aa polypeptide. TaZFP15 contains two C2H2 zinc finger domains and each one has a conserved motif QALGGH. The typical L-box, generally identified in the C2H2 type transcription factors, has also been found in TaZFP15. Phylogenetic analysis suggested that TaZFP15 shares high similarities with rice ZFP15 （GenBank accession no. AY286473）, maize ZFP （GenBank accession no. NM_001159094） and a subset of other zinc-finger transcription factor genes in plant species. The expression of TaZFP15 was up-regulated by starved-Pi stress, showing a pattern to be gradually elevated along with the progression of the Pi-stress in a 23-h treatment regime. Similarly, the transcripts of TaZFP15 in roots were also induced by nitrogen deficiency, and abiotic stresses of drought and salinity. No responses of TaZFP15 were detected in roots to nutrition deficiencies of P, Zn, and Ca, and the external treatment of abscisic acid （ABA）. TaZFP15 could be specifically amplified in genome A, B, and D, and without variability in the sequences, suggesting that TaZFP15 has multi-copies in the homologous hexaploid species. Transgenic analysis in tobacco revealed that up-regulation of TaZFP15 could significantly improve plant dry mass accumulation via increasing the plant phosphorus acquisition capacity under Pi-deficiency condition. The results suggested that TaZFP15 is involved in mediation of signal transductions of diverse external stresses.
基金Supported by the National Key Technology Research and Development Program (2006BAD03A01)the Hi-Tech Research and Development Program of China (2007AA10Z106)the Key Program Project of Ministry of Education (104242).
文摘The Populus euphratica stress responsive zinc-finger protein gene PSTZ, which encodes a protein including typical Cys2/His2 zinc finger structure, was isolated by reverse transcription-polymerase chain reaction from P. euphratica. Northern hybridization revealed that its expression was induced under drought and salt stress conditions. To examine its function, cDNA of the PSTZ gene, driven by the cauliflower mosaic virus 35S promoter, was cloned into a plant expression vector pBin438 and introduced into tobacco plants. Transgenic tobacco showed an enhanced salt tolerance, suggesting that PSTZ may play a role in plant responsiveness to salt stress.
文摘Ikaros represents a zinc-finger protein family important for lymphocyte development and certain other physiological processes. The number of family members is large, with alternative splicing producing various additional isoforms from each of the five homologous genes in the family. The functional forms of Ikaros proteins could be even more diverse due to protein–protein interactions readily established between family members. Emerging evidence suggests that targeting Ikaros proteins is feasible and effective in therapeutic applications, although the exact roles of Ikaros proteins remain elusive within the intricate regulatory networks in which they are involved. In this review we collect existing knowledge as to the functions, regulatory pathways, and molecular mechanisms of this family of proteins in an attempt to gain a better understanding through the comparison of activities and interactions among family members.
基金Supported by High Technology Research and Development Program of China (863 Program, No. 2006AA100309)
文摘An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.
基金supported by the National Natural Science Foundation of China (Grant No. 31770804)CAMS Initiative for Innovative Medicine, China (Grant No. 2016-I2M-1–009)the IMM Basic Research Fund, China (Grant No. 2014ZD03)
文摘CCCTC-binding factor(CTCF) is a zinc-finger protein, serving an important part in the genome architecture as well as some biochemical processes. Over 70,000 CTCF binding DNA sites have been detected genome-wide, and most anchors of chromatin loops are demarcated with the CTCF binding.Various protein or RNA molecules interact with DNA-bound CTCF to conduct different biological functions, and potentially the interfaces between CTCF and its cofactors can be targets for drug development. Here we identify the effective region of CTCF in DNA recognition, which defines the exposed CTCF surface feature for the interaction of cofactors. While the zinc-finger region contributes the most in DNA association, its binding affinity varies based on different DNA sequences. To investigate the effectiveness of individual zinc-fingers, the key residues are mutated to inactivate the DNA binding ability, while the finger configuration and the spacing between fingers are preserved. The strategy is proved to be successful, while clear differences are observed in the DNA binding affinities among the 11 finger mutants and the result is consistent to previous studies in general. With the help of inactivated finger mutants, we identify the ineffective fingers and the dominant effective fingers, which form distinctive patterns on different DNA targets.
基金supported by the Taishan Scholar Seed Industry Plan,Shandong Province,China
文摘RING is a really interesting new gene which plays important regulatory roles in many developmental processes as well as in plant-environment interactions. In the present report, the Zm RHCP1 gene encoding a putative RING-HC protein was isolated from maize and characterized. The Zm RHCP1 protein contained 310 amino acid residues with a conserved RINGHC zinc-finger motif and two transmembrane（TM） domains. Zm RHCP1 was expressed ubiquitously in various organs（root, stem, leaf, seedling, immature ear, and tassel）, but its transcript levels were higher in vegetative organs than in reproductive organs. Moreover, the expression pattern of Zm RHCP1 in brace roots indicated that Zm RHCP1 functions in brace root initiation. In addition, Zm RHCP1 expression was regulated by abiotic stresses. The expression results suggested that Zm RHCP1 plays important roles in brace root development and abiotic stress responses. The findings of the present study provide important information to help us understand the function of Zm RHCP1 in maize.
基金supported in part by Grants(to G.G.)from National Natural Science Foundation of China(Grant Nos.30470092 and 30530020)National Basic Research Program of China(973 Program)(Grant No.2006CB504302)of China.
文摘The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses,including murine leukemia virus,Sindbis virus and Ebola virus,by targeting the viral mRNAs for degradation.ZAP directly binds to the target viral mRNA and recruits the cellular RNA degradation machinery to degrade the RNA.No significant sequence similarity or obvious common motifs have been found in the so far identified target viral mRNAs.The minimum length of the target sequence is about 500 nt long.Short workable ZAP-binding RNAs should facilitate further studies on the ZAP-RNA interaction and characterization of such RNAs may provide some insights into the underlying mechanism.In this study,we used the SELEX method to isolate ZAP-binding RNA aptamers.After 21 rounds of selection,ZAP-binding aptamers were isolated.Sequence analysis revealed that they are G-rich RNAs with predicted stem-loop structures containing conserved“GGGUGG”and“GAGGG”motifs in the loop region.Insertion of the aptamer sequence into a luciferase reporter failed to render the reporter sensitive to ZAP.However,overexpression of the aptamers modestly but significantly reduced ZAP’s antiviral activity.Substitution of the conserved motifs of the aptamers significantly impaired their ZAP-binding ability and ZAP-antagonizing activity,suggesting that the RNA sequence is important for specific interaction between ZAP and the target RNA.The aptamers identified in this report should provide useful tools to further investigate the details of the interaction between ZAP and the target RNAs.
文摘Well-established targeted technologies to engi- neer genomes such as zinc-finger nuclease-based editing （ZFN）, transcription activator-like effector nuclease-based editing （TALEN）, and clustered regularly interspaced short palindromic repeats and associated protein system-based editing （CRISPR/Cas） are proving to advance basic and applied research in numerous plant species. Compared with systems using ZFNs and TALENs, the most recently developed CRISPR/Cas system is more efficient due to its use of an RNA-guided nuclease to generate double-strand DNA breaks. To accelerate the applications of these technologies, we provide here a detailed overview of these systems, highlight the strengths and weaknesses of each, summarize research advances made with these technologies in model and crop plants, and discuss their applications in plant functional genomics. Such targeted approaches for genetically modifying plants will benefit agricultural production in the future.
基金Work on this topic in the authors’laboratories is supported by grants from:the Strategic Priority Research Program of the Chinese Academy of Sciences(number XDA01020106)the Ministry of Science and Technology of China 973 program(2011CB965200)+2 种基金the National Natural Science Foundation of China(81261130317)to MAEthe Bureau of Science,Technology and Information of Guangzhou Municipality(2012 J5100040)to MAE and JFgrants 2010U1-E00811-5 and ZNGI-2011-010 from the Guangzhou Municipality and the Chinese Academy of Sciences,respectively,to LL.
文摘Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platformscontributing to redefine the boundaries of modern biological research. They are composed of a non-specificcleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications byinducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases havebeen employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively.This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies,biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbitsusing transcription activator-like effector nucleases, and a perspective of the field.
基金supported by Shanghai Municipal Health Commission(Grant/Award No.20194Y0050).
文摘Background:LncRNA DLX6-AS1 has been uncovered to exert effects on various cancers.Nevertheless,the impacts of DLX6-AS1 on endometrial cancer(EC)development remained obscure.The study explored the influence of DLX6-AS1 on EC progression via the microRNA(miR)-374a-3p/zinc-finger protein(ZFX)axis.Methods:EC cell lines were collected and DLX6-AS1,miR-374a-3p,and ZFX levels in EC cell lines were detected.The EC cells were transfected with DLX6-AS1,miR-374a-3p,and ZFX constructs to examine the biological functions of EC cells.The xenograft model was established for detecting tumor growth.Rescue experiments were conducted to verify the interaction of DLX6-AS1,miR-374a-3p,and ZFX in EC cells.Results:DLX6-AS1 and ZFX levels were elevated,while miR-374a-3p exhibited a reduced level in EC cells.Silencing DLX6-AS1 and elevated miR-374a-3p expressions repressed the biological activities of EC cells.Reduced DLX6-AS1 repressed tumor development.MiR-374a-3p silencing reversed the impacts of DLX6-AS1 silencing,while ZFX overexpression abrogated the impacts of miR-374a-3p elevation on EC cell growth.Mechanically,DLX6-AS1 was found to bind to miR-374a-3p,and miR-374a-3p targeted ZFX.Conclusion:DLX6-AS1 depletion restricts the malignant phenotype of EC cells.The study might provide novel therapeutic biomarkers for EC treatment.
基金Supported by grants from Medical Science and Technology Research Fund of Guangdong Province(No.WSTJJ20111110440104197405153780)the Dean Fund of Nanfang Hospital,Southern Medical University(No.2012B015)
文摘The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.
基金supported by the grant to Guangxia Gao from National Science Foundation of China(Grant No.81030030)by the grant toGuifang Chen from National Science Foundation(Grant No.30800053).
文摘The zinc-finger antiviral protein(ZAP)is a host factor that specifically inhibits the replication of certain viruses by eliminating viral mRNAs in the cytoplasm.In previous studies,we demonstrated that ZAP directly binds to the viral mRNAs and recruits the RNA exosome to degrade the target RNA.In this article,we provide evidence that a DEXH box RNA helicase,DHX30,is required for optimal antiviral activity of ZAP.Pull-down and co-immunoprecipitation assays demonstrated that DHX30 and ZAP interacted with each other via their N terminal domains.Downregulation of DHX30 with shRNAs reduced ZAP’s antiviral activity.These data implicate that DHX30 is a cellular factor involved in the antiviral function of ZAP.
基金supported by the National Natural Science Foundation of China(Grant No.31971919)the National Key Program for Research and Development of China(Grant No.2017YFD0100202)+1 种基金the Project Sponsored by Natural Science Foundation of Chongqing,China(Grant No.cstc2020jcyjjqX0020)Chongqing Graduate Research and Innovation Project funding in China(Grant No.CYS20123)。
文摘Inflorescence structure of rice,including the number and length of branches,and the density of the spikelet,can greatly affect the number of grains per panicle,which is one of the key factors in yield compositions.Here we identified five allelic mutants sb1-1/2/3/4/5 that related to branch development of rice.In these mutants,the branch meristem fate was prolonged sharply,resulting in delay of transition from branches to spikelets,and then increased the numbers of branches and spikelets per panicle.SB1 encodes a nuclear RING-like domain protein of SHI/LRP/SRS family and strongly expressed in branch meristems.The results of protein interaction and chromatin immunoprecipitation further suggested that SB1 directly repressed the expression of DEP1,TAW1,MOC1 and IPA1 by interacting with a co-repressor complex to affect acetylation level of histone H3 on target regions.Thus,we proposed that SB1 is a transcription repressor of branch meristem activity by widely and negatively regulating a series of genes that maintain branch meristem fate.
文摘Functional biological research has benefited tremendously by analyses of the phenotypes of mutant organisms which can be generated through targeted mutation of genes.In Drosophila,compared with random mutagenesis methods gene targeting has gained its popularity because it can introduce any desired mutation into a gene of interest.However,applications of gene targeting have been limited because the targeting efficiency varies with different genes,and the time and labor of targeting procedure are intensive.Nevertheless,improvement of gene targeting and development of its variant technologies have received much attention of scientists.Here we review recent progress that has been made in expanding the applications of gene targeting,which include the ϕC31 integration system and zinc-finger nucleases induced gene targeting,and new strategies that generate more efficient and reliable gene targeting.
文摘Despite relative effectiveness of current hepatitis B therapies,there is still no curative agents available.The new emerging approaches hold promise to achieve cure and loss of hepatitis B surface antigen.Studies or clinical trials investigating new therapies remain small and either focus on patients with low viral load and without hepatotoxic injury or patients with hepatitis D co-infection,which makes it challenging to assess their effectiveness and side effect profile in hepatitis B population.
文摘Sp1/Krüppel样因子(Sp1-like and Krüppel-like factors,Sp1/KLFs)是一组与真核细胞转录调控密切相关的锌指蛋白。Sp1/KLFs的羧基末端高度保守,含有3个串联的Cys2His2锌指结构,用于结合DNA;其氨基末端在不同的家族成员间存在较大差异,主要是通过结合辅助因子发挥转录调控作用。Sp1/KLFs的表达具有组织、细胞分布以及发育时期的特异性,它们通过调控多种富含GC或CACCC的启动子的基因的表达,参与细胞增殖、分化、凋亡和肿瘤发生、发展等多种生理、病理过程。文章综述了Sp1/KLFs的结构特征、作用机制及生物学功能。
