Synthesis, debugging, and effects of synthetic chromosome consolidation: synVI and beyond

Leslie A. Mitchell; Ann Wang; Giovanni Stracquadanio; Zheng Kuang; Xuya Wang; Kun Yang; Sarah Richardson; J. Andrew Martin; Yu Zhao; Roy Walker; Yisha Luo; Hongjiu Dai; Kang Dong; Zuojian Tang; Yanling Yang; Yizhi Cai; Adriana Heguy; Beatrix Ueberheide; David Fenyö; Junbiao Dai; Joel S. Bader; Jef D. Boeke

doi:10.1126/science.aaf4831

Synthesis, debugging, and effects of synthetic chromosome consolidation

synVI and beyond

Leslie A. Mitchell, Ann Wang, Giovanni Stracquadanio, Zheng Kuang, Xuya Wang, Kun Yang, Sarah Richardson, J. Andrew Martin, Yu Zhao, Roy Walker, Yisha Luo, Hongjiu Dai, Kang Dong, Zuojian Tang, Yanling Yang, Yizhi Cai, Adriana Heguy, Beatrix Ueberheide, David Fenyö, Junbiao Dai & 2 others Joel S. Bader, Jef D. Boeke^*

^*Corresponding author for this work

Research output: Contribution to journal › Article

97 Citations (Scopus)

Abstract

We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within PRE4 (YFR050C), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis, associated with alteration of the HIS2 transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome, no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.

Original language	English
Article number	eaaf4831
Pages (from-to)	1-11
Number of pages	12
Journal	Science
Volume	355
Issue number	6329
DOIs	https://doi.org/10.1126/science.aaf4831
Publication status	Published - 10 Mar 2017
Externally published	Yes

Bibliographical note

Plus 1 page summary.

Access to Document

10.1126/science.aaf4831

Link to publication in Scopus

Fingerprint Dive into the research topics of 'Synthesis, debugging, and effects of synthetic chromosome consolidation: synVI and beyond'. Together they form a unique fingerprint.

Cite this

Mitchell, Leslie A. ; Wang, Ann ; Stracquadanio, Giovanni ; Kuang, Zheng ; Wang, Xuya ; Yang, Kun ; Richardson, Sarah ; Martin, J. Andrew ; Zhao, Yu ; Walker, Roy ; Luo, Yisha ; Dai, Hongjiu ; Dong, Kang ; Tang, Zuojian ; Yang, Yanling ; Cai, Yizhi ; Heguy, Adriana ; Ueberheide, Beatrix ; Fenyö, David ; Dai, Junbiao ; Bader, Joel S. ; Boeke, Jef D. / Synthesis, debugging, and effects of synthetic chromosome consolidation : synVI and beyond. In: Science. 2017 ; Vol. 355, No. 6329. pp. 1-11.

@article{fa03cae4c06d4c79aebd54d117ea5bc5,

title = "Synthesis, debugging, and effects of synthetic chromosome consolidation: synVI and beyond",

abstract = "We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within PRE4 (YFR050C), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another {"}bug{"} was discovered through proteomic analysis, associated with alteration of the HIS2 transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6{\%} of the genome, no major global changes were detected in the poly-synthetic strain {"}omics{"} analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.",

author = "Mitchell, {Leslie A.} and Ann Wang and Giovanni Stracquadanio and Zheng Kuang and Xuya Wang and Kun Yang and Sarah Richardson and Martin, {J. Andrew} and Yu Zhao and Roy Walker and Yisha Luo and Hongjiu Dai and Kang Dong and Zuojian Tang and Yanling Yang and Yizhi Cai and Adriana Heguy and Beatrix Ueberheide and David Feny{\"o} and Junbiao Dai and Bader, {Joel S.} and Boeke, {Jef D.}",

note = "Plus 1 page summary.",

year = "2017",

month = "3",

day = "10",

doi = "10.1126/science.aaf4831",

language = "English",

volume = "355",

pages = "1--11",

journal = "Science (New York, N.Y.)",

issn = "0036-8075",

publisher = "American Association for the Advancement of Science",

number = "6329",

}

Mitchell, LA, Wang, A, Stracquadanio, G, Kuang, Z, Wang, X, Yang, K, Richardson, S, Martin, JA, Zhao, Y, Walker, R, Luo, Y, Dai, H, Dong, K, Tang, Z, Yang, Y, Cai, Y, Heguy, A, Ueberheide, B, Fenyö, D, Dai, J, Bader, JS & Boeke, JD 2017, 'Synthesis, debugging, and effects of synthetic chromosome consolidation: synVI and beyond', Science, vol. 355, no. 6329, eaaf4831, pp. 1-11. https://doi.org/10.1126/science.aaf4831

Synthesis, debugging, and effects of synthetic chromosome consolidation : synVI and beyond. / Mitchell, Leslie A.; Wang, Ann; Stracquadanio, Giovanni; Kuang, Zheng; Wang, Xuya; Yang, Kun; Richardson, Sarah; Martin, J. Andrew; Zhao, Yu; Walker, Roy; Luo, Yisha; Dai, Hongjiu; Dong, Kang; Tang, Zuojian; Yang, Yanling; Cai, Yizhi; Heguy, Adriana; Ueberheide, Beatrix; Fenyö, David; Dai, Junbiao; Bader, Joel S.; Boeke, Jef D.

In: Science, Vol. 355, No. 6329, eaaf4831, 10.03.2017, p. 1-11.

Research output: Contribution to journal › Article

TY - JOUR

T1 - Synthesis, debugging, and effects of synthetic chromosome consolidation

T2 - synVI and beyond

AU - Mitchell, Leslie A.

AU - Wang, Ann

AU - Stracquadanio, Giovanni

AU - Kuang, Zheng

AU - Wang, Xuya

AU - Yang, Kun

AU - Richardson, Sarah

AU - Martin, J. Andrew

AU - Zhao, Yu

AU - Walker, Roy

AU - Luo, Yisha

AU - Dai, Hongjiu

AU - Dong, Kang

AU - Tang, Zuojian

AU - Yang, Yanling

AU - Cai, Yizhi

AU - Heguy, Adriana

AU - Ueberheide, Beatrix

AU - Fenyö, David

AU - Dai, Junbiao

AU - Bader, Joel S.

AU - Boeke, Jef D.

N1 - Plus 1 page summary.

PY - 2017/3/10

Y1 - 2017/3/10

N2 - We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within PRE4 (YFR050C), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis, associated with alteration of the HIS2 transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome, no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.

AB - We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within PRE4 (YFR050C), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis, associated with alteration of the HIS2 transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome, no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.

UR - http://www.scopus.com/inward/record.url?scp=85014834845&partnerID=8YFLogxK

U2 - 10.1126/science.aaf4831

DO - 10.1126/science.aaf4831

M3 - Article

VL - 355

SP - 1

EP - 11

JO - Science (New York, N.Y.)

JF - Science (New York, N.Y.)

SN - 0036-8075

IS - 6329

M1 - eaaf4831

ER -