Bug mapping and fitness testing of chemically synthesized chromosome X

Science. 2017 Mar 10;355(6329):eaaf4706. doi: 10.1126/science.aaf4706.


Debugging a genome sequence is imperative for successfully building a synthetic genome. As part of the effort to build a designer eukaryotic genome, yeast synthetic chromosome X (synX), designed as 707,459 base pairs, was synthesized chemically. SynX exhibited good fitness under a wide variety of conditions. A highly efficient mapping strategy called pooled PCRTag mapping (PoPM), which can be generalized to any watermarked synthetic chromosome, was developed to identify genetic alterations that affect cell fitness ("bugs"). A series of bugs were corrected that included a large region bearing complex amplifications, a growth defect mapping to a recoded sequence in FIP1, and a loxPsym site affecting promoter function of ATP2 PoPM is a powerful tool for synthetic yeast genome debugging and an efficient strategy for phenotype-genotype mapping.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chromosomes, Artificial, Yeast / chemistry*
  • Chromosomes, Artificial, Yeast / genetics*
  • Gene Duplication
  • Genetic Fitness
  • Genome, Fungal*
  • High-Throughput Nucleotide Sequencing / methods*
  • Physical Chromosome Mapping / methods*
  • Saccharomyces cerevisiae / genetics*
  • Synthetic Biology