Clonal tracking using embedded viral barcoding and high-throughput sequencing

Nat Protoc. 2020 Apr;15(4):1436-1458. doi: 10.1038/s41596-019-0290-z. Epub 2020 Mar 4.

Abstract

Embedded viral barcoding in combination with high-throughput sequencing is a powerful technology with which to track single-cell clones. It can provide clonal-level insights into cellular proliferation, development, differentiation, migration, and treatment efficacy. Here, we present a detailed protocol for a viral barcoding procedure that includes the creation of barcode libraries, the viral delivery of barcodes, the recovery of barcodes, and the computational analysis of barcode sequencing data. The entire procedure can be completed within a few weeks. This barcoding method requires cells to be susceptible to viral transduction. It provides high sensitivity and throughput, and enables precise quantification of cellular progeny. It is cost efficient and does not require any advanced skills. It can also be easily adapted to many types of applications, including both in vitro and in vivo experiments.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation / genetics
  • Cell Tracking / methods*
  • Clone Cells / cytology*
  • DNA / genetics
  • DNA Barcoding, Taxonomic / methods*
  • Genetic Vectors / genetics
  • HEK293 Cells
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Lentivirus / genetics
  • Mice

Substances

  • DNA