Digital microfluidics-based digital counting of single-cell copy number variation (dd-scCNV Seq)

作者全名："Yu, Xiyuan; Ruan, Weidong; Lin, Fanghe; Qian, Weizhou; Zou, Yuan; Liu, Yilong; Su, Rui; Niu, Qi; Ruan, Qingyu; Lin, Wei; Zhu, Zhi; Zhang, Huimin; Yang, Chaoyong"

作者地址："[Yu, Xiyuan; Ruan, Weidong; Lin, Fanghe; Qian, Weizhou; Liu, Yilong; Niu, Qi; Ruan, Qingyu; Zhu, Zhi; Yang, Chaoyong] Xiamen Univ, Coll Chem & Chem Engn, Dept Chem Biol, Key Lab Spectrochem Anal & Instrumentat,Minist Edu, Xiamen 361005, Peoples R China; [Zou, Yuan] Chongqing Med Univ, Coll Lab Med, Key Lab Clin Lab Diagnost, Minist Educ, Chongqing 400016, Peoples R China; [Su, Rui] Xiamen Univ, Affiliated Hosp 1, Dept Hematol, Xiamen 361005, Peoples R China; [Lin, Wei; Zhang, Huimin; Yang, Chaoyong] Innovat Lab Sci & Technol Energy Mat Fujian Prov, Xiamen 361005, Peoples R China"

通信作者："Yang, CY (通讯作者)，Xiamen Univ, Coll Chem & Chem Engn, Dept Chem Biol, Key Lab Spectrochem Anal & Instrumentat,Minist Edu, Xiamen 361005, Peoples R China.; Zhang, HM; Yang, CY (通讯作者)，Innovat Lab Sci & Technol Energy Mat Fujian Prov, Xiamen 361005, Peoples R China."

来源：PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA

ESI学科分类：Multidisciplinary

WOS号：WOS:001038273900002

JCR分区：Q1

影响因子：9.4

年份：2023

卷号：120

期号：20

开始页：　

结束页：　

文献类型：Article

关键词：single-cell analysis; digital counting; transposon insertion; copy number variation; genome sequencing

摘要："Single-cell copy number variations (CNVs), major dynamic changes in humans, result in differential levels of gene expression and account for adaptive traits or underlying disease. Single-cell sequencing is needed to reveal these CNVs but has been hindered by single-cell whole-genome amplification (scWGA) bias, leading to inaccurate gene copy number counting. In addition, most of the current scWGA methods are labor intensive, time-consuming, and expensive with limited wide application. Here, we report a unique single-cell whole-genome library preparation approach based on digital microfluidics for digital counting of single-cell Copy Number Variation (dd-scCNV Seq). dd-scCNV Seq directly fragments the original single-cell DNA and uses these fragments as templates for amplification. These reduplicative fragments can be filtered computationally to generate the original partitioned unique identified fragments, thereby enabling digital counting of copy number variation. dd-scCNV Seq showed an increase in uniformity in the single-molecule data, leading to more accurate CNV patterns compared to other methods with low-depth sequencing. Benefiting from digital microfluidics, dd-scCNV Seq allows automated liquid handling, precise single-cell isolation, and high-efficiency and low-cost genome library preparation. dd-scCNV Seq will accelerate biological discovery by enabling accurate profiling of copy number variations at single-cell resolution."

基金机构："National Natural Science Foundation of China [21904085, 21927806, 21735004, 21974113]; National Key Ramp;D Program of China [2019YFA0905800, 2021YFA0909400]; Fundamental Research Funds for the Central Universities [20720210001, 20720220005]"

基金资助正文："ACKNOWLEDGMENTS. We thank the National Natural Science Foundation of China (21904085, 21927806, 21735004, and 21974113) , the National Key R&D Program of China (2019YFA0905800, 2021YFA0909400) , and the Fundamental Research Funds for the Central Universities (20720210001, 20720220005) for their financial support."