SeqVerify: An accessible analysis tool for cell line genomic integrity, contamination, and gene editing outcomes

Stem Cell Reports. 2024 Oct 8;19(10):1505-1515. doi: 10.1016/j.stemcr.2024.08.004. Epub 2024 Sep 12.

Abstract

Over the last decade, advances in genome editing and pluripotent stem cell (PSC) culture have let researchers generate edited PSC lines to study a wide variety of biological questions. However, abnormalities in cell lines such as aneuploidy, mutations, on-target and off-target editing errors, and microbial contamination can arise during PSC culture or due to undesired editing outcomes. The ongoing decline of next-generation sequencing prices has made whole-genome sequencing (WGS) a promising option for detecting these abnormalities. However, this approach has been held back by a lack of easily usable data analysis software. Here, we present SeqVerify, a computational pipeline designed to take raw WGS data and a list of intended genome edits, and verify that the edits are present and that there are no abnormalities. We anticipate that SeqVerify will be a useful tool for researchers generating edited PSCs, and more broadly, for cell line quality control in general.

Keywords: SeqVerify; aneuploidy; genome editing; microbial contamination; pluripotent stem cell; quality control; single-nucleotide polymorphisms; software; stem cell; whole-genome sequencing.

MeSH terms

  • Animals
  • CRISPR-Cas Systems
  • Cell Line
  • Computational Biology / methods
  • Gene Editing* / methods
  • Genomics / methods
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism
  • Software*
  • Whole Genome Sequencing / methods