Engineered yeast genomes accurately assembled from pure and mixed samples
Abstract
Yeast whole genome sequencing (WGS) lacks end-to-end workflows that identify genetic engineering. Here we present Prymetime, a tool that assembles yeast plasmids and chromosomes and annotates genetic engineering sequences. It is a hybrid workflow—it uses short and long reads as inputs to perform separate linear and circular assembly steps. This structure is necessary to accurately resolve genetic engineering sequences in plasmids and the genome. We show this by assembling diverse engineered yeasts, in some cases revealing unintended deletions and integrations. Furthermore, the resulting whole genomes are high quality, although the underlying assembly software does not consistently resolve highly repetitive genome features. Finally, we assemble plasmids and genome integrations from metagenomic sequencing, even with 1 engineered cell in 1000. This work is a blueprint for building WGS workflows and establishes WGS-based identification of yeast genetic engineering.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 05, 2021
- Source ID
- 10.1038/s41467-021-21656-9
Entities
People
- Aaron Adler
- Bryan A Bartley
- Celeste B. Marsan
- Eric M Young
- Joseph H. Collins
- Kevin Keating
- Marina Çomo
- Nicholas Roehner
- Shravani Balaji
- Tom M. Mitchell
- Trent R. Jones
- Zachary J. Newlon
Organizations
- Intelligence Advanced Research Projects Activity