Antibody upstream sequence diversity and its biological implications revealed by repertoire sequencing

查看全文 作　　者：Yan [1,2,3,4,5]Zhu;Xiujia [1,2,3,4,5]Yang;Cuiyu [1,2]Ma;Haipei [3]Tang;Qilong [3]Wang;Junjie [1,2]Guan;Wenxi [1,2]Xie;Sen [1,2]Chen;Yuan [3]Chen;Minhui [1,6,7]Wang;Chunhong [1,3]Lan;Deqiang [8]Sun;Lai [9]Wei;Caijun [10]Sun;Xueqing [4,11]Yu;Zhenhai [1,2,3,4,5]Zhang 机构地区：[1]State Key Laboratory of Organ Failure Research,National Clinical Research Center for Kidney Disease,Division of Nephrology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;[2]Department of Bioinformatics,School of Basic Medical Sciences,Southern Medical University,Guangzhou 510515,China;[3]Center for Precision Medicine,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;[4]Guangdong-Hong Kong Joint Laboratory on Immunological and Genetic Kidney Diseases,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;[5]Key Laboratory of Mental Health of the Ministry of Education,Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence,Southern Medical University,Guangzhou 510515,China;[6]Department of Nephrology,Hainan Affiliated Hospital of Hainan Medical College,Haikou 570311,China;[7]Department of Nephrology,Hainan General Hospital,Haikou 570311,China;[8]Department of Center Laboratory,The Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China;[9]State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510060,China;[10]School of Public Health,Sun Yat-sen University,Shenzhen 510006,China;[11]Division of Nephrology,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China 出　　处：《Journal of Genetics and Genomics》索引2021年第48卷第10期,共10页 基　　金：supported by the National Natural Science Foundation of China(NSFC)(31771479 to Z.Z.);NSFC Projects of International Cooperation and Exchanges of NSFC(61661146004 to Z.Z.);the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(2017BT01S131 to Z.Z.);Guangdong-Hong Kong-Macao-Joint Labs Program from Guangdong Science and Technology(2019B121205005 to X.Y.)。 摘　　要：The sequence upstream of the antibody variable region(antibody upstream sequence[AUS])consists of a 5′untranslated region(5′UTR)and a preceding leader region.The sequence variations in AUS affect antibody engineering and PCR based antibody quantification and may also be implicated in mRNA transcription and translation.However,the diversity of AUSs remains elusive.Using 5′rapid amplification of cDNA ends and high-throughput antibody repertoire sequencing technique,we acquired full-length AUSs for human,rhesus macaque,cynomolgus macaque,mouse,and rat.We designed a bioinformatics pipeline and identified 3307 unique AUSs,corresponding to 3026 and 1457 unique sequences for 5′UTR and leader region,respectively.Comparative analysis indicated that 928(63.69%)leader sequences are novel relative to those recorded in the international ImMunoGeneTics information system.Evolutionarily,leader sequences are more conserved than 5′UTR and seem to coevolve with their downstream V genes.Besides,single-nucleotide polymorphisms are position dependent for leader regions and may contribute to the functional reversal of the downstream V genes.Finally,the AUGs in AUSs were found to have little impact on gene expression.Taken together,our findings can facilitate primer design for capturing antibodies efficiently and provide a valuable resource for antibody engineering and molecule-level antibody studies. 关 键 词：Antibody upstream sequences 5′UTR Leader sequences Antibody repertoire sequencing Antibody repertoire