Single-cell whole-genome analyses by Linear Amplification via Transposon Insertion (LIANTI).

Science
Authors
Keywords
Abstract

Single-cell genomics is important for biology and medicine. However, current whole-genome amplification (WGA) methods are limited by low accuracy of copy-number variation (CNV) detection and low amplification fidelity. Here we report an improved single-cell WGA method, Linear Amplification via Transposon Insertion (LIANTI), which outperforms existing methods, enabling micro-CNV detection with kilobase resolution. This allowed direct observation of stochastic firing of DNA replication origins, which differs from cell to cell. We also show that the predominant cytosine-to-thymine mutations observed in single-cell genomics often arise from the artifact of cytosine deamination upon cell lysis. However, identifying single-nucleotide variations (SNVs) can be accomplished by sequencing kindred cells. We determined the spectrum of SNVs in a single human cell after ultraviolet radiation, revealing their nonrandom genome-wide distribution.

Year of Publication
2017
Journal
Science
Volume
356
Issue
6334
Pages
189-194
Date Published
2017 04 14
ISSN
1095-9203
DOI
10.1126/science.aak9787
PubMed ID
28408603
PubMed Central ID
PMC5538131
Links
Grant list
DP1 CA186693 / CA / NCI NIH HHS / United States
R33 CA174560 / CA / NCI NIH HHS / United States