PgmNr Y3148: Synthesis, debugging and consolidation of synthetic chromosomes in yeast: synVI and beyond.
Authors:
Leslie A. Mitchell
1,2 ; Ann Wang
3,4 ; Giovanni Stracquadanio
3,5 ; Zheng Kuang
1,2 ; Xuya Wang
1,2 ; Sarah Richardson
3,5 ; J. Andrew Martin
1,2 ; Roy Walker
6 ; Yisha Luo
6 ; Hongjiu Dai
7 ; Kang Dong
7 ; Zuojian Tang
8 ; Yizhi Cai
6 ; David Fenyo
8 ; Junbiao Dai
4 ; Joel S. Bader
3,4 ; Jef D. Boeke
1,2
Institutes
1) Department of Biochemistry and Molecular Pharmacology, New York University Langone School of Medicine, New York City, NY 10016, USA; 2) Institute for Systems Genetics, New York University Langone School of Medicine, New York City, NY 10016, USA; 3) High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; 4) Key laboratory for Industrial Biocatalysis (Ministry of Education), Key laboratory of Bioinformatics (Ministry of Education), Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China; 5) Department of Biomedical Engineering and Institute of Genetic Medicine, Whiting School of Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; 6) Center for Synthetic and Systems Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH9 3JL; 7) GenScript, Piscataway, NJ 08854, USA; 8) Center for Health Informatics and Bioinformatics, New York University Langone School of Medicine, New York City, NY 10016, USA. Keyword: Genomics
Abstract: The Synthetic Yeast Genome project, Sc2.0, aims to build a designer genome to power growth of the yeast Saccharomyces cerevisiae. Here we describe the design, rapid assembly, and characterization of Sc2.0 chromosome VI, dubbed synVI. Growth defects observed in the synVI strain were mapped to the synthetic chromosome, including one that affected mitochondrial behavior. We mapped the mitochondrial defect to a series of synonymous coding sequence changes in the essential gene PRE4, encoding a proteasomal subunit; coding changes reduced Pre4 protein accumulation by about half. Completing Sc2.0 necessitates a method to backcross synthetic chromosome strains to wild type and to combine multiple synthetic chromosomes into a single strain. We developed a method to rapidly cross synthetic chromosomes to any strain of interest without sequence alteration and built a poly-synthetic strain encoding synIII, synVI, and synIXR. This work sets the stage for completion of the world’s first synthetic, designer eukaryotic genome.
Yeast Database Genetic Index
1. gene symbol: PRE4; systematic name: YRF050C