Title:
METHOD FOR AMPLIFYING NUCLEIC ACID USING SOLID-PHASE CARRIER
Document Type and Number:
WIPO Patent Application WO/2021/006353
Kind Code:
A1
Abstract:
This method for amplifying a nucleic acid using a solid-phase carrier includes capturing a target nucleic acid including mRNA on a solid-phase carrier and synthesizing a complementary strand on the solid phase, and, after decomposing/removing the unreacted target nucleic acid for capture on the solid phase by exonuclease processing, performing homopolymer addition by mRNA decomposition and TdT reaction in the presence of a chain-terminating nucleotide triphosphate. Through this method, cDNA can be stably amplified with high efficiency even from a minute sample, regardless of an excess quantity of enzyme relative to the DNA substrate on the solid phase, excess reaction time, or differences in reagent lots. Through this amplification method, it is possible to expand the range of application of a technique in which analysis using DNA-labeled antibodies or other specific binding molecules labeled with an oligonucleic acid and analysis of transcripts are performed simultaneously.
Inventors:
MATSUSHIMA KOUJI (JP)
UEHA SATOSHI (JP)
SHICHINO SHIGEYUKI (JP)
ITO SATORU (JP)
AOKI HIROYASU (JP)
UEHA SATOSHI (JP)
SHICHINO SHIGEYUKI (JP)
ITO SATORU (JP)
AOKI HIROYASU (JP)
Application Number:
PCT/JP2020/027123
Publication Date:
January 14, 2021
Filing Date:
July 10, 2020
Export Citation:
Assignee:
UNIV TOKYO SCIENCE FOUND (JP)
International Classes:
C12N15/10; C12Q1/6806; C12Q1/686; C12Q1/6869
Domestic Patent References:
| WO2015166768A1 | 2015-11-05 | |||
| WO2013145431A1 | 2013-10-03 | |||
| WO2015027135A1 | 2015-02-26 | |||
| WO2015200893A2 | 2015-12-30 | |||
| WO2013145431A1 | 2013-10-03 | |||
| WO2016040476A1 | 2016-03-17 | |||
| WO2015166768A1 | 2015-11-05 |
Foreign References:
| US20160312276A1 | 2016-10-27 | |||
| US20180251825A1 | 2018-09-06 | |||
| JP2016533187A | 2016-10-27 | |||
| JP2017522867A | 2017-08-17 | |||
| US20180258500A1 | 2018-09-13 |
Other References:
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AOKI HIROYASU, UEHA SATOSHI, SHICHINO SHIGEYUKI, OGIWARA HARU, HASHIMOTO SHIN-ICHI, KAKIMI KAZUHIRO, ITO SATORU, MATSUSHIMA KOUJI: "TCR Repertoire Analysis Reveals Mobilization of Novel CD8+ T Cell Clones Into the Cancer-Immunity Cycle Following Anti-CD4 Antibody Administration", FRONTIERS IN IMMUNOLOGY, vol. 9, 1 January 2018 (2018-01-01), pages 3185, XP055785297, DOI: 10.3389/fimmu.2018.03185
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ZARETSKY JMGARCIA-DIAZ A. SHIN DS ET AL.: "Mutations Associated with Acquired Resistance to PD-1 Blockade in Melanoma", N ENGL J MED, vol. 375, 2016, pages 819 - 2
AOKI H. ET AL.: "TCR Repertoire Analysis Reveals Mobilization of Novel CD8+ T Cell Clones Into the Cancer-Immunity Cycle Following Anti-CD4 Antibody Administration", FRONT IMMUNOL, vol. 9, no. 3185, January 2019 (2019-01-01)
PICELLI S. ET AL.: "Full-length RNA-seq from single cells using Smart-seq2", NAT PROTOC, vol. 9, no. 1, January 2014 (2014-01-01), pages 171 - 81, XP002742134, DOI: 10.1038/nprot.2014.006
SINGLE-CELL TRANSCRIPTOMICS OF 20 MOUSE ORGANS CREATES A TABULA MURIS, NATURE, vol. 562, no. 7727, October 2018 (2018-10-01), pages 367 - 372
MATSUNAGA HGOTO MARIKAWA K ET AL.: "A highly sensitive and accurate gene expression analysis by sequencing (''bead-seq'') for a single cell", ANAL BIOCHEM, vol. 471, 2015, pages 9 - 16
HUAN HUANGMARI GOTOHIROYUKI TSUNODALIZHOU SUNKIYOMI TANIGUCHIHIROKO MATSUNAGAHIDEKI KAMBARA: "Non-biased and efficient global amplification of a single-cell cDNA library", NUCLEIC ACID RESEARCH, 2013
SASAGAWA YNIKAIDO IHAYASHI TDANNO HUNO KDIMAI TUEDA HR: "Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity", GENOME BIOL, 2013
YOHEI SASAGAWAHIROKI DANNOHITOMI TAKADAMASASHI EBISAWAKAORI TANAKATETSUTARO HAYASHIAKIRA KURISAKIITOSHI NIKAIDO: "Quartz-Seq2: a high-throughput single-cell RNA-sequencing method that effectively uses limited sequence reads", GENOME BIOL, 2018
MACOSKO EZ ET AL.: "Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets", CELL, vol. 161, no. 5, 2015, pages 1202 - 1214, XP055586617, DOI: 10.1016/j.cell.2015.05.002
GIERAHN TM: "Seq-Well: portable, low-cost RNA sequencing of single cells at high throughput", NAT METHODS, vol. 14, no. 4, 2017, pages 395 - 398
HASHIMOTO ET AL., SCIENTIFIC REPORTS, vol. 7, no. I, 27 October 2017 (2017-10-27), pages 14225
MERRIFIELD, BIOCHEMISTRY, vol. 3, 1964, pages 1385 - 1390
See also references of EP 3998338A4
MARLON STOECKIUS, HAFEMEISTER CHRISTOPH, STEPHENSON WILLIAM, HOUCK-LOOMIS BRIAN, CHATTOPADHYAY PRATIP K, SWERDLOW HAROLD, SATIJA R: "Simultaneous epitope and transcriptome measurement in single cells", NATURE METHODS, NATURE PUB. GROUP, NEW YORK, vol. 14, no. 9, 1 September 2017 (2017-09-01), New York, pages 865 - 868, XP055547724, ISSN: 1548-7091, DOI: 10.1038/nmeth.4380
AOKI HIROYASU, UEHA SATOSHI, SHICHINO SHIGEYUKI, OGIWARA HARU, HASHIMOTO SHIN-ICHI, KAKIMI KAZUHIRO, ITO SATORU, MATSUSHIMA KOUJI: "TCR Repertoire Analysis Reveals Mobilization of Novel CD8+ T Cell Clones Into the Cancer-Immunity Cycle Following Anti-CD4 Antibody Administration", FRONTIERS IN IMMUNOLOGY, vol. 9, 1 January 2018 (2018-01-01), pages 3185, XP055785297, DOI: 10.3389/fimmu.2018.03185
REGEV A. ET AL.: "The Human Cell Atlas", ELIFE, 2017, pages e27041
ZARETSKY JMGARCIA-DIAZ A. SHIN DS ET AL.: "Mutations Associated with Acquired Resistance to PD-1 Blockade in Melanoma", N ENGL J MED, vol. 375, 2016, pages 819 - 2
AOKI H. ET AL.: "TCR Repertoire Analysis Reveals Mobilization of Novel CD8+ T Cell Clones Into the Cancer-Immunity Cycle Following Anti-CD4 Antibody Administration", FRONT IMMUNOL, vol. 9, no. 3185, January 2019 (2019-01-01)
PICELLI S. ET AL.: "Full-length RNA-seq from single cells using Smart-seq2", NAT PROTOC, vol. 9, no. 1, January 2014 (2014-01-01), pages 171 - 81, XP002742134, DOI: 10.1038/nprot.2014.006
SINGLE-CELL TRANSCRIPTOMICS OF 20 MOUSE ORGANS CREATES A TABULA MURIS, NATURE, vol. 562, no. 7727, October 2018 (2018-10-01), pages 367 - 372
MATSUNAGA HGOTO MARIKAWA K ET AL.: "A highly sensitive and accurate gene expression analysis by sequencing (''bead-seq'') for a single cell", ANAL BIOCHEM, vol. 471, 2015, pages 9 - 16
HUAN HUANGMARI GOTOHIROYUKI TSUNODALIZHOU SUNKIYOMI TANIGUCHIHIROKO MATSUNAGAHIDEKI KAMBARA: "Non-biased and efficient global amplification of a single-cell cDNA library", NUCLEIC ACID RESEARCH, 2013
SASAGAWA YNIKAIDO IHAYASHI TDANNO HUNO KDIMAI TUEDA HR: "Quartz-Seq: a highly reproducible and sensitive single-cell RNA sequencing method, reveals non-genetic gene-expression heterogeneity", GENOME BIOL, 2013
YOHEI SASAGAWAHIROKI DANNOHITOMI TAKADAMASASHI EBISAWAKAORI TANAKATETSUTARO HAYASHIAKIRA KURISAKIITOSHI NIKAIDO: "Quartz-Seq2: a high-throughput single-cell RNA-sequencing method that effectively uses limited sequence reads", GENOME BIOL, 2018
MACOSKO EZ ET AL.: "Highly Parallel Genome-wide Expression Profiling of Individual Cells Using Nanoliter Droplets", CELL, vol. 161, no. 5, 2015, pages 1202 - 1214, XP055586617, DOI: 10.1016/j.cell.2015.05.002
GIERAHN TM: "Seq-Well: portable, low-cost RNA sequencing of single cells at high throughput", NAT METHODS, vol. 14, no. 4, 2017, pages 395 - 398
HASHIMOTO ET AL., SCIENTIFIC REPORTS, vol. 7, no. I, 27 October 2017 (2017-10-27), pages 14225
MERRIFIELD, BIOCHEMISTRY, vol. 3, 1964, pages 1385 - 1390
See also references of EP 3998338A4
Attorney, Agent or Firm:
TANIGAWA AND PARTNERS, PATENT FIRM (JP)
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