{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T20:50:45Z","timestamp":1790628645874,"version":"4.1.0"},"reference-count":42,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,7,14]],"date-time":"2017-07-14T00:00:00Z","timestamp":1499990400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,7,14]],"date-time":"2017-07-14T00:00:00Z","timestamp":1499990400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Inverted terminal repeats (ITRs) of the adeno-associated virus (AAV) are essential for rescue, replication, packaging, and integration of the viral genome. While ITR mutations have been identified in previous reports, we designed a new truncated ITR lacking the B-B\u2019 and C-C\u2019 regions named as ITR\u0394BC and investigated its effects on viral genome replication, packaging, and expression of recombinant AAV (rAAV). The packaging ability was compared between ITR\u0394BC rAAV and wild-type (wt) ITR rAAV. Our results showed the productivity of ITR\u0394BC rAAV was reduced 4-fold, which is consistent with the 8-fold decrease in the replication of viral genomic DNA of ITR\u0394BC rAAV compared with wt ITR rAAV. Surprisingly, transgene expression was significantly higher for ITR\u0394BC rAAV. A preliminary exploration of the underlying mechanisms was carried out by inhibiting and degrading the ataxia telangiectasia mutated (ATM) protein and the Mre11 complex (MRN), respectively, since the rAAV expression was inhibited by the ATM and\/or MRN through <jats:italic>cis<\/jats:italic> interaction or binding with wt ITRs. We demonstrated that the inhibitory effects were weakened on ITR\u0394BC rAAV expression. This study suggests deletion in ITR can affect the transgene expression of AAV, which provides a new way to improve the AAV expression through ITRs modification.<\/jats:p>","DOI":"10.1038\/s41598-017-04054-4","type":"journal-article","created":{"date-parts":[[2017,7,10]],"date-time":"2017-07-10T10:55:07Z","timestamp":1499684107000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Deletion of the B-B\u2019 and C-C\u2019 regions of inverted terminal repeats reduces rAAV productivity but increases transgene expression"],"prefix":"10.1038","volume":"7","author":[{"given":"Qingzhang","family":"Zhou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenhong","family":"Tian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunguo","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhonghui","family":"Lian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyan","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaobing","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,7,14]]},"reference":[{"key":"4054_CR1","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1128\/jvi.45.2.555-564.1983","volume":"45","author":"A Srivastava","year":"1983","unstructured":"Srivastava, A., Lusby, E. W. & Berns, K. I. Nucleotide sequence and organization of the adeno-associated virus 2 genome. J Virol \n                           45, 555\u2013564 (1983).","journal-title":"J Virol"},{"key":"4054_CR2","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1038\/nrg2988","volume":"12","author":"F Mingozzi","year":"2011","unstructured":"Mingozzi, F. & High, K. A. Therapeutic in vivo gene transfer for genetic disease using AAV: progress and challenges. Nature reviews. Genetics \n                           12, 341\u2013355, doi:10.1038\/nrg2988 (2011).","journal-title":"Nature reviews. Genetics"},{"key":"4054_CR3","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1128\/jvi.34.2.402-409.1980","volume":"34","author":"E Lusby","year":"1980","unstructured":"Lusby, E., Fife, K. H. & Berns, K. I. Nucleotide sequence of the inverted terminal repetition in adeno-associated virus DNA. J Virol \n                           34, 402\u2013409 (1980).","journal-title":"J Virol"},{"key":"4054_CR4","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1016\/0092-8674(83)90342-2","volume":"33","author":"RJ Samulski","year":"1983","unstructured":"Samulski, R. J., Srivastava, A., Berns, K. I. & Muzyczka, N. Rescue of adeno-associated virus from recombinant plasmids: gene correction within the terminal repeats of AAV. Cell \n                           33, 135\u2013143 (1983).","journal-title":"Cell"},{"key":"4054_CR5","doi-asserted-by":"publisher","first-page":"2077","DOI":"10.1073\/pnas.79.6.2077","volume":"79","author":"RJ Samulski","year":"1982","unstructured":"Samulski, R. J., Berns, K. I., Tan, M. & Muzyczka, N. Cloning of adeno-associated virus into pBR322: rescue of intact virus from the recombinant plasmid in human cells. Proceedings of the National Academy of Sciences of the United States of America \n                           79, 2077\u20132081 (1982).","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"4054_CR6","doi-asserted-by":"crossref","first-page":"1128","DOI":"10.1128\/jvi.68.2.1128-1138.1994","volume":"68","author":"TH Ni","year":"1994","unstructured":"Ni, T. H., Zhou, X., McCarty, D. M., Zolotukhin, I. & Muzyczka, N. In vitro replication of adeno-associated virus DNA. J Virol \n                           68, 1128\u20131138 (1994).","journal-title":"J Virol"},{"key":"4054_CR7","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1038\/sj.mt.6300376","volume":"16","author":"L Zhong","year":"2008","unstructured":"Zhong, L. et al. Single-polarity recombinant adeno-associated virus 2 vector-mediated transgene expression in vitro and in vivo: mechanism of transduction. Molecular therapy: the journal of the American Society of Gene Therapy \n                           16, 290\u2013295, doi:10.1038\/sj.mt.6300376 (2008).","journal-title":"Molecular therapy: the journal of the American Society of Gene Therapy"},{"key":"4054_CR8","doi-asserted-by":"publisher","first-page":"2105","DOI":"10.1038\/sj.gt.3302133","volume":"10","author":"Z Wang","year":"2003","unstructured":"Wang, Z. et al. Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo. Gene therapy \n                           10, 2105\u20132111, doi:10.1038\/sj.gt.3302133 (2003).","journal-title":"Gene therapy"},{"key":"4054_CR9","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1146\/annurev.genet.37.110801.143717","volume":"38","author":"DM McCarty","year":"2004","unstructured":"McCarty, D. M., Young, S. M. Jr. & Samulski, R. J. Integration of adeno-associated virus (AAV) and recombinant AAV vectors. Annual review of genetics \n                           38, 819\u2013845, doi:10.1146\/annurev.genet.37.110801.143717 (2004).","journal-title":"Annual review of genetics"},{"key":"4054_CR10","doi-asserted-by":"crossref","first-page":"1668","DOI":"10.1128\/jvi.70.3.1668-1677.1996","volume":"70","author":"XS Wang","year":"1996","unstructured":"Wang, X. S., Ponnazhagan, S. & Srivastava, A. Rescue and replication of adeno-associated virus type 2 as well as vector DNA sequences from recombinant plasmids containing deletions in the viral inverted terminal repeats: selective encapsidation of viral genomes in progeny virions. J Virol \n                           70, 1668\u20131677 (1996).","journal-title":"J Virol"},{"key":"4054_CR11","doi-asserted-by":"crossref","first-page":"7448","DOI":"10.1128\/jvi.68.11.7448-7457.1994","volume":"68","author":"JA Chiorini","year":"1994","unstructured":"Chiorini, J. A. et al. Sequence requirements for stable binding and function of Rep68 on the adeno-associated virus type 2 inverted terminal repeats. J Virol \n                           68, 7448\u20137457 (1994).","journal-title":"J Virol"},{"key":"4054_CR12","doi-asserted-by":"publisher","first-page":"5887","DOI":"10.1128\/JVI.00254-08","volume":"82","author":"D Grimm","year":"2008","unstructured":"Grimm, D. et al. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol \n                           82, 5887\u20135911, doi:10.1128\/JVI.00254-08 (2008).","journal-title":"J Virol"},{"key":"4054_CR13","doi-asserted-by":"crossref","first-page":"2224","DOI":"10.1128\/JVI.72.3.2224-2232.1998","volume":"72","author":"X Xiao","year":"1998","unstructured":"Xiao, X., Li, J. & Samulski, R. J. Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus. J Virol \n                           72, 2224\u20132232 (1998).","journal-title":"J Virol"},{"key":"4054_CR14","doi-asserted-by":"publisher","first-page":"485","DOI":"10.1007\/BF03187263","volume":"46","author":"X Wu","year":"2001","unstructured":"Wu, X. et al. A novel method for purification of recombinant adenoassociated virus vectors on a large scale. Chinese Science Bulletin \n                           46, 485\u2013489 (2001).","journal-title":"Chinese Science Bulletin"},{"key":"4054_CR15","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1089\/hgtb.2012.090","volume":"23","author":"Y Wang","year":"2012","unstructured":"Wang, Y. et al. Limitations of encapsidation of recombinant self-complementary adeno-associated viral genomes in different serotype capsids and their quantitation. Human gene therapy methods \n                           23, 225\u2013233, doi:10.1089\/hgtb.2012.090 (2012).","journal-title":"Human gene therapy methods"},{"key":"4054_CR16","doi-asserted-by":"publisher","first-page":"11480","DOI":"10.1128\/JVI.77.21.11480-11490.2003","volume":"77","author":"P Ward","year":"2003","unstructured":"Ward, P., Elias, P. & Linden, R. M. Rescue of the adeno-associated virus genome from a plasmid vector: evidence for rescue by replication. J Virol \n                           77, 11480\u201311490 (2003).","journal-title":"J Virol"},{"key":"4054_CR17","doi-asserted-by":"publisher","first-page":"e29551","DOI":"10.1371\/journal.pone.0029551","volume":"7","author":"W Tian","year":"2012","unstructured":"Tian, W. et al. High-throughput functional microRNAs profiling by recombinant AAV-based microRNA sensor arrays. PloS one \n                           7, e29551, doi:10.1371\/journal.pone.0029551 (2012).","journal-title":"PloS one"},{"key":"4054_CR18","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1016\/j.ymthe.2004.10.016","volume":"11","author":"BA Tannous","year":"2005","unstructured":"Tannous, B. A., Kim, D. E., Fernandez, J. L., Weissleder, R. & Breakefield, X. O. Codon-optimized Gaussia luciferase cDNA for mammalian gene expression in culture and in vivo. Molecular therapy: the journal of the American Society of Gene Therapy \n                           11, 435\u2013443, doi:10.1016\/j.ymthe.2004.10.016 (2005).","journal-title":"Molecular therapy: the journal of the American Society of Gene Therapy"},{"key":"4054_CR19","doi-asserted-by":"publisher","first-page":"171","DOI":"10.1038\/nmeth.1177","volume":"5","author":"T Wurdinger","year":"2008","unstructured":"Wurdinger, T. et al. A secreted luciferase for ex vivo monitoring of in vivo processes. Nature methods \n                           5, 171\u2013173, doi:10.1038\/nmeth.1177 (2008).","journal-title":"Nature methods"},{"key":"4054_CR20","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1006\/viro.1999.0137","volume":"268","author":"S Sanlioglu","year":"2000","unstructured":"Sanlioglu, S., Benson, P. & Engelhardt, J. F. Loss of ATM function enhances recombinant adeno-associated virus transduction and integration through pathways similar to UV irradiation. Virology \n                           268, 68\u201378, doi:10.1006\/viro.1999.0137 (2000).","journal-title":"Virology"},{"key":"4054_CR21","doi-asserted-by":"publisher","first-page":"686","DOI":"10.1038\/gt.2012.86","volume":"20","author":"MP Cataldi","year":"2013","unstructured":"Cataldi, M. P. & McCarty, D. M. Hairpin-end conformation of adeno-associated virus genome determines interactions with DNA-repair pathways. Gene therapy \n                           20, 686\u2013693, doi:10.1038\/gt.2012.86 (2013).","journal-title":"Gene therapy"},{"key":"4054_CR22","doi-asserted-by":"publisher","first-page":"8673","DOI":"10.1128\/JVI.00641-10","volume":"84","author":"MP Cataldi","year":"2010","unstructured":"Cataldi, M. P. & McCarty, D. M. Differential effects of DNA double-strand break repair pathways on single-strand and self-complementary adeno-associated virus vector genomes. J Virol \n                           84, 8673\u20138682, doi:10.1128\/JVI.00641-10 (2010).","journal-title":"J Virol"},{"key":"4054_CR23","doi-asserted-by":"publisher","first-page":"12936","DOI":"10.1128\/JVI.01523-07","volume":"81","author":"RA Schwartz","year":"2007","unstructured":"Schwartz, R. A. et al. The Mre11\/Rad50\/Nbs1 complex limits adeno-associated virus transduction and replication. J Virol \n                           81, 12936\u201312945, doi:10.1128\/JVI.01523-07 (2007).","journal-title":"J Virol"},{"key":"4054_CR24","doi-asserted-by":"publisher","first-page":"181","DOI":"10.1128\/JVI.01990-14","volume":"89","author":"TB Lentz","year":"2015","unstructured":"Lentz, T. B. & Samulski, R. J. Insight into the mechanism of inhibition of adeno-associated virus by the Mre11\/Rad50\/Nbs1 complex. J Virol \n                           89, 181\u2013194, doi:10.1128\/JVI.01990-14 (2015).","journal-title":"J Virol"},{"key":"4054_CR25","doi-asserted-by":"publisher","first-page":"14004","DOI":"10.1128\/JVI.79.22.14004-14016.2005","volume":"79","author":"Y Liu","year":"2005","unstructured":"Liu, Y., Shevchenko, A., Shevchenko, A. & Berk, A. J. Adenovirus exploits the cellular aggresome response to accelerate inactivation of the MRN complex. J Virol \n                           79, 14004\u201314016, doi:10.1128\/JVI.79.22.14004-14016.2005 (2005).","journal-title":"J Virol"},{"key":"4054_CR26","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/S0014-5793(96)01149-0","volume":"397","author":"BM Bishop","year":"1996","unstructured":"Bishop, B. M., Santin, A. D., Quirk, J. G. & Hermonat, P. L. Role of the terminal repeat GAGC trimer, the major Rep78 binding site, in adeno-associated virus DNA replication. FEBS letters \n                           397, 97\u2013100 (1996).","journal-title":"FEBS letters"},{"key":"4054_CR27","doi-asserted-by":"publisher","first-page":"e1000985","DOI":"10.1371\/journal.ppat.1000985","volume":"6","author":"D Huser","year":"2010","unstructured":"Huser, D. et al. Integration preferences of wildtype AAV-2 for consensus rep-binding sites at numerous loci in the human genome. PLoS pathogens \n                           6, e1000985, doi:10.1371\/journal.ppat.1000985 (2010).","journal-title":"PLoS pathogens"},{"key":"4054_CR28","doi-asserted-by":"crossref","first-page":"4988","DOI":"10.1128\/jvi.68.8.4988-4997.1994","volume":"68","author":"DM McCarty","year":"1994","unstructured":"McCarty, D. M. et al. Identification of linear DNA sequences that specifically bind the adeno-associated virus Rep protein. J Virol \n                           68, 4988\u20134997 (1994).","journal-title":"J Virol"},{"key":"4054_CR29","doi-asserted-by":"crossref","first-page":"3542","DOI":"10.1128\/jvi.69.6.3542-3548.1995","volume":"69","author":"RS Wonderling","year":"1995","unstructured":"Wonderling, R. S., Kyostio, S. R. & Owens, R. A. A maltose-binding protein\/adeno-associated virus Rep68 fusion protein has DNA-RNA helicase and ATPase activities. J Virol \n                           69, 3542\u20133548 (1995).","journal-title":"J Virol"},{"key":"4054_CR30","doi-asserted-by":"crossref","first-page":"1542","DOI":"10.1128\/jvi.70.3.1542-1553.1996","volume":"70","author":"JH Ryan","year":"1996","unstructured":"Ryan, J. H., Zolotukhin, S. & Muzyczka, N. Sequence requirements for binding of Rep68 to the adeno-associated virus terminal repeats. J Virol \n                           70, 1542\u20131553 (1996).","journal-title":"J Virol"},{"key":"4054_CR31","doi-asserted-by":"publisher","first-page":"7762","DOI":"10.1128\/JVI.74.17.7762-7771.2000","volume":"74","author":"JR Brister","year":"2000","unstructured":"Brister, J. R. & Muzyczka, N. Mechanism of Rep-mediated adeno-associated virus origin nicking. J Virol \n                           74, 7762\u20137771 (2000).","journal-title":"J Virol"},{"key":"4054_CR32","doi-asserted-by":"publisher","first-page":"494","DOI":"10.1038\/sj.mt.6300397","volume":"16","author":"X Zhou","year":"2008","unstructured":"Zhou, X. et al. Adeno-associated virus of a single-polarity DNA genome is capable of transduction in vivo. Molecular therapy: the journal of the American Society of Gene Therapy \n                           16, 494\u2013499, doi:10.1038\/sj.mt.6300397 (2008).","journal-title":"Molecular therapy: the journal of the American Society of Gene Therapy"},{"key":"4054_CR33","doi-asserted-by":"publisher","first-page":"12792","DOI":"10.1128\/JVI.76.24.12792-12802.2002","volume":"76","author":"S Musatov","year":"2002","unstructured":"Musatov, S., Roberts, J., Pfaff, D. & Kaplitt, M. A cis-acting element that directs circular adeno-associated virus replication and packaging. J Virol \n                           76, 12792\u201312802 (2002).","journal-title":"J Virol"},{"key":"4054_CR34","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1016\/0042-6822(88)90502-8","volume":"166","author":"RA Bohenzky","year":"1988","unstructured":"Bohenzky, R. A., LeFebvre, R. B. & Berns, K. I. Sequence and symmetry requirements within the internal palindromic sequences of the adeno-associated virus terminal repeat. Virology \n                           166, 316\u2013327 (1988).","journal-title":"Virology"},{"key":"4054_CR35","doi-asserted-by":"publisher","first-page":"4080","DOI":"10.1128\/JVI.75.9.4080-4090.2001","volume":"75","author":"J Hansen","year":"2001","unstructured":"Hansen, J., Qing, K. & Srivastava, A. Adeno-associated virus type 2-mediated gene transfer: altered endocytic processing enhances transduction efficiency in murine fibroblasts. J Virol \n                           75, 4080\u20134090, doi:10.1128\/JVI.75.9.4080-4090.2001 (2001).","journal-title":"J Virol"},{"key":"4054_CR36","doi-asserted-by":"crossref","first-page":"8568","DOI":"10.1128\/JVI.72.11.8568-8577.1998","volume":"72","author":"D Duan","year":"1998","unstructured":"Duan, D. et al. Circular intermediates of recombinant adeno-associated virus have defined structural characteristics responsible for long-term episomal persistence in muscle tissue. J Virol \n                           72, 8568\u20138577 (1998).","journal-title":"J Virol"},{"key":"4054_CR37","doi-asserted-by":"publisher","first-page":"952","DOI":"10.1128\/JVI.02581-14","volume":"89","author":"C Ling","year":"2015","unstructured":"Ling, C. et al. Enhanced transgene expression from recombinant single-stranded D-sequence-substituted adeno-associated virus vectors in human cell lines in vitro and in murine hepatocytes in vivo. J Virol \n                           89, 952\u2013961, doi:10.1128\/JVI.02581-14 (2015).","journal-title":"J Virol"},{"key":"4054_CR38","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1073\/pnas.56.1.86","volume":"56","author":"JA Rose","year":"1966","unstructured":"Rose, J. A., Hoggan, M. D. & Shatkin, A. J. Nucleic acid from an adeno-associated virus: chemical and physical studies. Proceedings of the National Academy of Sciences of the United States of America \n                           56, 86\u201392 (1966).","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"4054_CR39","doi-asserted-by":"publisher","first-page":"1955","DOI":"10.1172\/JCI34316","volume":"118","author":"M Allocca","year":"2008","unstructured":"Allocca, M. et al. Serotype-dependent packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice. The Journal of clinical investigation \n                           118, 1955\u20131964, doi:10.1172\/JCI34316 (2008).","journal-title":"The Journal of clinical investigation"},{"key":"4054_CR40","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1038\/mt.2009.256","volume":"18","author":"Y Lai","year":"2010","unstructured":"Lai, Y., Yue, Y. & Duan, D. Evidence for the failure of adeno-associated virus serotype 5 to package a viral genome > or\u2009=\u20098.2 kb. Molecular therapy: the journal of the American Society of Gene Therapy \n                           18, 75\u201379, doi:10.1038\/mt.2009.256 (2010).","journal-title":"Molecular therapy: the journal of the American Society of Gene Therapy"},{"key":"4054_CR41","doi-asserted-by":"publisher","first-page":"1412","DOI":"10.1038\/nprot.2006.207","volume":"1","author":"JC Grieger","year":"2006","unstructured":"Grieger, J. C., Choi, V. W. & Samulski, R. J. Production and characterization of adeno-associated viral vectors. Nature protocols \n                           1, 1412\u20131428, doi:10.1038\/nprot.2006.207 (2006).","journal-title":"Nature protocols"},{"key":"4054_CR42","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1016\/S0166-0934(02)00138-6","volume":"106","author":"UP Rohr","year":"2002","unstructured":"Rohr, U. P. et al. Fast and reliable titration of recombinant adeno-associated virus type-2 using quantitative real-time PCR. Journal of virological methods \n                           106, 81\u201388 (2002).","journal-title":"Journal of virological methods"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04054-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04054-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04054-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,23]],"date-time":"2022-12-23T06:39:03Z","timestamp":1671777543000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04054-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,7,14]]},"references-count":42,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["4054"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-04054-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,7,14]]},"assertion":[{"value":"7 December 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"9 May 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 July 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"Mr. Chunguo Liu and Dr. Xiaoyan Dong are employees of Beijing FivePlus Molecular Medicine Institute, Ltd. Dr. Xiaoyan Dong is the stockholder of the company. None of the other authors declare conflicts of interest.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"5432"}}