Open AccessClonal tracking using embedded viral barcoding and high-throughput sequencing
Author(s) -
Charles Bramlett,
Jie Du,
Anogalska,
Jiya Eerdeng,
Jorge L. Contreras,
Rong Lu
Publication year - 2020
Publication title -
nature protocols
Language(s) - English
Resource type - Journals
eISSN - 1754-2189
pISSN - 1750-2799
DOI - 10.1038/s41596-019-0290-z
Subject(s) - computational biology , dna sequencing , throughput , biology , dna barcoding , evolutionary biology , genetics , computer science , gene , telecommunications , wireless
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.