Accurate SNV Detection in Single Cells by Transposon-based Whole-genome Amplification of Complementary Strands

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2021 Feb 17

PMID 33593904

Citations 38

Authors

Dong Xing

Longzhi Tan

Chi-Han Chang

Heng Li

X Sunney Xie

Affiliations

Soon will be listed here.

Abstract

Single-nucleotide variants (SNVs), pertinent to aging and disease, occur sporadically in the human genome, hence necessitating single-cell measurements. However, detection of single-cell SNVs suffers from false positives (FPs) due to intracellular single-stranded DNA damage and the process of whole-genome amplification (WGA). Here, we report a single-cell WGA method termed multiplexed end-tagging amplification of complementary strands (META-CS), which eliminates nearly all FPs by virtue of DNA complementarity, and achieved the highest accuracy thus far. We validated META-CS by sequencing kindred cells and human sperm, and applied it to other human tissues. Investigation of mature single human neurons revealed increasing SNVs with age and potentially unrepaired strand-specific oxidative guanine damage. We determined SNV frequencies along the genome in differentiated single human blood cells, and identified cell type-dependent mutational patterns for major types of lymphocytes.

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