Background The primary differentially methylated regions(DMRs) which are maternally hypermethylated serve as imprinting control regions(ICRs) that drive monoallelic gene expression, and these ICRs have been investigat...Background The primary differentially methylated regions(DMRs) which are maternally hypermethylated serve as imprinting control regions(ICRs) that drive monoallelic gene expression, and these ICRs have been investigated due to their implications in mammalian development. Although a subset of genes has been identified as imprinted, in-depth comparative approach needs to be developed for identification of species-specific imprinted genes. Here, we examined DNA methylation status and allelic expression at the KBTBD6 locus across species and tissues and explored potential mechanisms of imprinting.Results Using whole-genome bisulfite sequencing and RNA-sequencing on parthenogenetic and normal porcine embryos, we identified a maternally hypermethylated DMR between the embryos at the KBTBD6 promoter Cp G island and paternal monoallelic expression of KBTBD6. Also, in analyzed domesticated mammals but not in humans, non-human primates and mice, the KBTBD6 promoter Cp G islands were methylated in oocytes and/or allelically methyl-ated in tissues, and monoallelic KBTBD6 expression was observed, indicating livestock-specific imprinting. Further analysis revealed that these Cp G islands were embedded within transcripts in porcine and bovine oocytes which coexisted with an active transcription mark and DNA methylation, implying the presence of transcription-dependent imprinting.Conclusions In this study, our comparative approach revealed an imprinted expression of the KBTBD6 gene in domesticated mammals, but not in humans, non-human primates, and mice which implicates species-specific evolution of genomic imprinting.展开更多
Copper is a microelement with important physiological functions in the body.However,the excess copper ion(Cu^(2+))may cause severe health problems,such as hair cell apoptosis and the resultant hearing loss.Therefore,t...Copper is a microelement with important physiological functions in the body.However,the excess copper ion(Cu^(2+))may cause severe health problems,such as hair cell apoptosis and the resultant hearing loss.Therefore,the assay of Cu^(2+)is important.We integrate ionic imprinting technology(IIT)and structurally colored hydrogel beads to prepare chitosan-based ionically imprinted hydrogel beads(IIHBs)as a low-cost and high-specificity platform for Cu^(2+)detection.The IIHBs have a macroporous microstructure,uniform size,vivid structural color,and magnetic responsiveness.When incubated in solution,IIHBs recognize Cu^(2+)and exhibit a reflective peak change,thereby achieving label-free detection.In addition,benefiting from the IIT,the IIHBs display good specificity and selectivity and have an imprinting factor of 19.14 at 100μmol·L^(-1).These features indicated that the developed IIHBs are promising candidates for Cu^(2+)detection,particularly for the prevention of hearing loss.展开更多
利用溶剂热法制备纳米Fe 3 O 4,并将其分散在正硅酸乙酯(TEOS)水解液中,在表面沉积一层SiO_(2).用甲基丙烯酰氧基丙基三甲氧基硅烷(MPS)对SiO_(2)-Fe 3 O 4表面改性,得到MPS-SiO_(2)-Fe 3 O 4.将甲基丙烯酸(MAA)与己烯雌酚(DES)以摩尔比...利用溶剂热法制备纳米Fe 3 O 4,并将其分散在正硅酸乙酯(TEOS)水解液中,在表面沉积一层SiO_(2).用甲基丙烯酰氧基丙基三甲氧基硅烷(MPS)对SiO_(2)-Fe 3 O 4表面改性,得到MPS-SiO_(2)-Fe 3 O 4.将甲基丙烯酸(MAA)与己烯雌酚(DES)以摩尔比4∶1加入乙腈溶剂中,进行12 h自组装,再加入MPS-SiO_(2)-Fe 3 O 4、乙二醇二甲基丙烯酸酯(EGDMA)和偶氮二异丁腈(AIBN),进行超声分散30 min,65℃条件下机械搅拌,反应24 h,制得DES磁性分子印迹聚合物(MMIP).采用透射电镜、振动样品磁强计和吸附试验等方法进行表征分析.结果表明:MMIP的饱和磁化强度为268 kA/m,无磁滞现象,矫顽力为0,表现出超顺磁性,在磁铁作用下17 s就可与溶液分离;室温下,MMIP对DES的静态最大吸附量为7.1 mg/g,动态吸附时MMIP在60 min时可以达到静态最大吸附量的90%以上;MMIP对DES的印迹因子为3.70,因而对DES有较大的识别能力;MMIP可重复使用6次以上,具有良好的再生循环性能.展开更多
基金partially supported by the United States Department of Agriculture National Institute of Food and Agriculture Hatch Grant (Project No.OHO01304)。
文摘Background The primary differentially methylated regions(DMRs) which are maternally hypermethylated serve as imprinting control regions(ICRs) that drive monoallelic gene expression, and these ICRs have been investigated due to their implications in mammalian development. Although a subset of genes has been identified as imprinted, in-depth comparative approach needs to be developed for identification of species-specific imprinted genes. Here, we examined DNA methylation status and allelic expression at the KBTBD6 locus across species and tissues and explored potential mechanisms of imprinting.Results Using whole-genome bisulfite sequencing and RNA-sequencing on parthenogenetic and normal porcine embryos, we identified a maternally hypermethylated DMR between the embryos at the KBTBD6 promoter Cp G island and paternal monoallelic expression of KBTBD6. Also, in analyzed domesticated mammals but not in humans, non-human primates and mice, the KBTBD6 promoter Cp G islands were methylated in oocytes and/or allelically methyl-ated in tissues, and monoallelic KBTBD6 expression was observed, indicating livestock-specific imprinting. Further analysis revealed that these Cp G islands were embedded within transcripts in porcine and bovine oocytes which coexisted with an active transcription mark and DNA methylation, implying the presence of transcription-dependent imprinting.Conclusions In this study, our comparative approach revealed an imprinted expression of the KBTBD6 gene in domesticated mammals, but not in humans, non-human primates, and mice which implicates species-specific evolution of genomic imprinting.
基金supported by grants from the National Key Research and Development Program of China(2021YFA1101300,2021YFA1101800,and 2020YFA0112503)the National Natural Science Foundation of China(82030029,81970882,92149304,and 22302231)+5 种基金the Science and Technology Department of Sichuan Province(2021YFS0371)the Guangdong Basic and Applied Basic Research Foundation(2023A1515011986)the Shenzhen Fundamental Research Program(JCYJ20190814093401920,JCYJ20210324125608022,JCYJ20190813152616459,and JCYJ20190808120405672)the Futian Healthcare Research Project(FTWS2022013 and FTWS2023080)the Open Research Fund of State Key Laboratory of Genetic Engineering,Fudan University(SKLGE-2104)the Fundamental Research Funds for the Central Universities,Sun Yat-sen University(23qnpy153)。
文摘Copper is a microelement with important physiological functions in the body.However,the excess copper ion(Cu^(2+))may cause severe health problems,such as hair cell apoptosis and the resultant hearing loss.Therefore,the assay of Cu^(2+)is important.We integrate ionic imprinting technology(IIT)and structurally colored hydrogel beads to prepare chitosan-based ionically imprinted hydrogel beads(IIHBs)as a low-cost and high-specificity platform for Cu^(2+)detection.The IIHBs have a macroporous microstructure,uniform size,vivid structural color,and magnetic responsiveness.When incubated in solution,IIHBs recognize Cu^(2+)and exhibit a reflective peak change,thereby achieving label-free detection.In addition,benefiting from the IIT,the IIHBs display good specificity and selectivity and have an imprinting factor of 19.14 at 100μmol·L^(-1).These features indicated that the developed IIHBs are promising candidates for Cu^(2+)detection,particularly for the prevention of hearing loss.
文摘利用溶剂热法制备纳米Fe 3 O 4,并将其分散在正硅酸乙酯(TEOS)水解液中,在表面沉积一层SiO_(2).用甲基丙烯酰氧基丙基三甲氧基硅烷(MPS)对SiO_(2)-Fe 3 O 4表面改性,得到MPS-SiO_(2)-Fe 3 O 4.将甲基丙烯酸(MAA)与己烯雌酚(DES)以摩尔比4∶1加入乙腈溶剂中,进行12 h自组装,再加入MPS-SiO_(2)-Fe 3 O 4、乙二醇二甲基丙烯酸酯(EGDMA)和偶氮二异丁腈(AIBN),进行超声分散30 min,65℃条件下机械搅拌,反应24 h,制得DES磁性分子印迹聚合物(MMIP).采用透射电镜、振动样品磁强计和吸附试验等方法进行表征分析.结果表明:MMIP的饱和磁化强度为268 kA/m,无磁滞现象,矫顽力为0,表现出超顺磁性,在磁铁作用下17 s就可与溶液分离;室温下,MMIP对DES的静态最大吸附量为7.1 mg/g,动态吸附时MMIP在60 min时可以达到静态最大吸附量的90%以上;MMIP对DES的印迹因子为3.70,因而对DES有较大的识别能力;MMIP可重复使用6次以上,具有良好的再生循环性能.