The emergence of single-cell genomic and transcriptomic sequencing accelerates the development of single-cell epigenomic technologies,providing an unprecedented opportunity for decoding cell fate decisions largely enc...The emergence of single-cell genomic and transcriptomic sequencing accelerates the development of single-cell epigenomic technologies,providing an unprecedented opportunity for decoding cell fate decisions largely encoded in the epigenome.Recent advances in single-cell multimodality epigenomic technologies facilitate directly interrogating the reg-ulatory relationship between multi-layer molecular information in the same cell.In this review,we discuss recent progress in development of single-cell multimodality epigenomic technologies and applications in elucidating cellular diversifications in development and diseases,with a focus on protein-DNA interactomics and regulatory links between epigenome and tran-scriptome.Further,we provide perspective on the future direction of single-cell multiomics tool development as well as challenges facing ahead.展开更多
目的构建和评价HPV16 E6、E7多表位DNA疫苗诱导的特异性CTL细胞应答及其对肿瘤生长的干预作用,从而揭示其作为候选HPV治疗性疫苗的潜能。方法首先通过IEDB网站中的MHC I Processing Predictions和MHC I Binding Predictions方法,分别预...目的构建和评价HPV16 E6、E7多表位DNA疫苗诱导的特异性CTL细胞应答及其对肿瘤生长的干预作用,从而揭示其作为候选HPV治疗性疫苗的潜能。方法首先通过IEDB网站中的MHC I Processing Predictions和MHC I Binding Predictions方法,分别预测人类HLA-A^(*)02:01、HLA-A^(*)11:01、HLA-A^(*)24:02和C57BL/6小鼠H-2b的限制性CTL表位,然后根据评分以及ELISPOT实验筛选出二者共同呈递的CTL表位,并将其构建成多表位DNA疫苗(pVAX1-10P)。从预防性和治疗性二个方面研究pVAX1-10P对小鼠移植TC-1异位癌的免疫干预作用,流式细胞术检测特异性CTL应答。结果获得10条可被人与鼠MHC分子共呈递的CTL表位,ELISPOT结果表明这10条CTL表位均能诱导小鼠淋巴细胞产生特异性免疫应答;由此构建的多表位DNA疫苗pVAX1-10P无论在预防性实验还是治疗性实验中,均能诱导特异性的细胞免疫并抑制肿瘤的生长。结论构建的HPV16 E6、E7多表位DNA疫苗pVAX1-10P能够诱导特异性CTL应答,显著抑制肿瘤生长,有望作为候选HPV治疗性DNA疫苗。展开更多
HL-60 cells have been induced with differentiation index 16 % by S-adenosyl-L-rnethionine (SAM) as inducer in the presence of optimum conceptration of 10 μmol/L. The methylation level of genorne DNA determined by HPL...HL-60 cells have been induced with differentiation index 16 % by S-adenosyl-L-rnethionine (SAM) as inducer in the presence of optimum conceptration of 10 μmol/L. The methylation level of genorne DNA determined by HPLC is increased during cell differentiation. When restriction endonuclease Hae Ⅲ, Sma I, Sal I, XhoI and Hind Ⅲ which are sensitive to 5-methylcytosine were used to cleave the genorne DNA, a resistance effect was found. The interaction between DNA and DNA binding proteins is changed by using gel retarding test.展开更多
目的探讨两种较为公认但序列不同的Wiskott-Aldrich综合征蛋白和富含脯氨酸同源物(Wiskott-Aldrich syndrome protein and SCAR homolog,WASH)的差异。方法使用免疫荧光、免疫共沉淀和激光微辐射等实验分析两种WASH在定位模式、与FAM21...目的探讨两种较为公认但序列不同的Wiskott-Aldrich综合征蛋白和富含脯氨酸同源物(Wiskott-Aldrich syndrome protein and SCAR homolog,WASH)的差异。方法使用免疫荧光、免疫共沉淀和激光微辐射等实验分析两种WASH在定位模式、与FAM21或Ku蛋白的相互作用、向DNA损伤位点募集速率和蛋白质稳定性方面的差异;比较多种生物中的WASH蛋白质序列和检测多种在生物和医学研究中常用的细胞中的WASH编码序列,分析两种人WASH序列的普遍性和保守性。结果两种WASH展现出类似的内体(endosome)定位模式。WASH468表现出与FAM21更强的相互作用,并且WASH468展现出更强的稳定性。WASH465表现出与Ku蛋白更强的相互作用,并且WASH465展现出向DNA损伤位点更强的募集。多种生物中的WASH序列与人WASH468序列的一致性明显高于WASH465,并且多种在生物和医学研究中常用的细胞中的WASH氨基酸序列均与WASH468一致。结论WASH468和WASH465的生物学特性存在差异,WASH468序列的普遍性和保守性明显高于WASH465,因此WASH468是更保守的人WASH序列。展开更多
基金supported by the National Key Research and Development Program of China(2021YFA1100101 and 2017YFA0103402)the National Natural Science Foundation of China(32025015 and 31771607)the Peking-Tsinghua Center for Life Sciences.
文摘The emergence of single-cell genomic and transcriptomic sequencing accelerates the development of single-cell epigenomic technologies,providing an unprecedented opportunity for decoding cell fate decisions largely encoded in the epigenome.Recent advances in single-cell multimodality epigenomic technologies facilitate directly interrogating the reg-ulatory relationship between multi-layer molecular information in the same cell.In this review,we discuss recent progress in development of single-cell multimodality epigenomic technologies and applications in elucidating cellular diversifications in development and diseases,with a focus on protein-DNA interactomics and regulatory links between epigenome and tran-scriptome.Further,we provide perspective on the future direction of single-cell multiomics tool development as well as challenges facing ahead.
文摘目的构建和评价HPV16 E6、E7多表位DNA疫苗诱导的特异性CTL细胞应答及其对肿瘤生长的干预作用,从而揭示其作为候选HPV治疗性疫苗的潜能。方法首先通过IEDB网站中的MHC I Processing Predictions和MHC I Binding Predictions方法,分别预测人类HLA-A^(*)02:01、HLA-A^(*)11:01、HLA-A^(*)24:02和C57BL/6小鼠H-2b的限制性CTL表位,然后根据评分以及ELISPOT实验筛选出二者共同呈递的CTL表位,并将其构建成多表位DNA疫苗(pVAX1-10P)。从预防性和治疗性二个方面研究pVAX1-10P对小鼠移植TC-1异位癌的免疫干预作用,流式细胞术检测特异性CTL应答。结果获得10条可被人与鼠MHC分子共呈递的CTL表位,ELISPOT结果表明这10条CTL表位均能诱导小鼠淋巴细胞产生特异性免疫应答;由此构建的多表位DNA疫苗pVAX1-10P无论在预防性实验还是治疗性实验中,均能诱导特异性的细胞免疫并抑制肿瘤的生长。结论构建的HPV16 E6、E7多表位DNA疫苗pVAX1-10P能够诱导特异性CTL应答,显著抑制肿瘤生长,有望作为候选HPV治疗性DNA疫苗。
文摘HL-60 cells have been induced with differentiation index 16 % by S-adenosyl-L-rnethionine (SAM) as inducer in the presence of optimum conceptration of 10 μmol/L. The methylation level of genorne DNA determined by HPLC is increased during cell differentiation. When restriction endonuclease Hae Ⅲ, Sma I, Sal I, XhoI and Hind Ⅲ which are sensitive to 5-methylcytosine were used to cleave the genorne DNA, a resistance effect was found. The interaction between DNA and DNA binding proteins is changed by using gel retarding test.
文摘目的探讨两种较为公认但序列不同的Wiskott-Aldrich综合征蛋白和富含脯氨酸同源物(Wiskott-Aldrich syndrome protein and SCAR homolog,WASH)的差异。方法使用免疫荧光、免疫共沉淀和激光微辐射等实验分析两种WASH在定位模式、与FAM21或Ku蛋白的相互作用、向DNA损伤位点募集速率和蛋白质稳定性方面的差异;比较多种生物中的WASH蛋白质序列和检测多种在生物和医学研究中常用的细胞中的WASH编码序列,分析两种人WASH序列的普遍性和保守性。结果两种WASH展现出类似的内体(endosome)定位模式。WASH468表现出与FAM21更强的相互作用,并且WASH468展现出更强的稳定性。WASH465表现出与Ku蛋白更强的相互作用,并且WASH465展现出向DNA损伤位点更强的募集。多种生物中的WASH序列与人WASH468序列的一致性明显高于WASH465,并且多种在生物和医学研究中常用的细胞中的WASH氨基酸序列均与WASH468一致。结论WASH468和WASH465的生物学特性存在差异,WASH468序列的普遍性和保守性明显高于WASH465,因此WASH468是更保守的人WASH序列。
