@article{850048ee5dff4a2cb7c27f7073f9ed54,
title = "Context-dependent neocentromere activity in synthetic yeast chromosome VIII",
abstract = "Pioneering advances in genome engineering, and specifically in genome writing, have revolutionized the field of synthetic biology, propelling us toward the creation of synthetic genomes. The Sc2.0 project aims to build the first fully synthetic eukaryotic organism by assembling the genome of Saccharomyces cerevisiae. With the completion of synthetic chromosome VIII (synVIII) described here, this goal is within reach. In addition to writing the yeast genome, we sought to manipulate an essential functional element: the point centromere. By relocating the native centromere sequence to various positions along chromosome VIII, we discovered that the minimal 118-bp CEN8 sequence is insufficient for conferring chromosomal stability at ectopic locations. Expanding the transplanted sequence to include a small segment (~500 bp) of the CDEIII-proximal pericentromere improved chromosome stability, demonstrating that minimal centromeres display context-dependent functionality ",
author = "Stephanie Lauer and Jingchuan Luo and Luciana Lazar-Stefanita and Weimin Zhang and McCulloch, {Laura H.} and Viola Fanfani and Evgenii Lobzaev and Haase, {Max A. B.} and Nicole Easo and Yu Zhao and Fangzhou Yu and Jitong Cai and {Build-A-Genome Class} and Bader, {Joel S.} and Giovanni Stracquadanio and Boeke, {Jef D.}",
note = "Funding Information: We thank Philip Hieter for helpful discussions on dicentric chromosomes. We thank Karen Yuen, Abby Mak, and Hin Ling for generously supplying strains and plasmids and for helpful discussions about the CEN8 minichromosome. We thank Megan Hogan, Raven Luther, Hannah Ashe, and Gwen Ellis from the Matthew Maurano lab, as well as Camila Coelho from the Boeke lab, for assistance with WGS. This work is part of the Sc2.0 project ( http://syntheticyeast.org/ ), supported by NSF grants MCB-1026068dslngs1 , MCB-1443299 , MCB-1616111 , and MCB-1921641 to J.D.B., as well as the UKRI EPSRC Fellowship ( EP/V033794/1 ) to G.S. Funding Information: We thank Philip Hieter for helpful discussions on dicentric chromosomes. We thank Karen Yuen, Abby Mak, and Hin Ling for generously supplying strains and plasmids and for helpful discussions about the CEN8 minichromosome. We thank Megan Hogan, Raven Luther, Hannah Ashe, and Gwen Ellis from the Matthew Maurano lab, as well as Camila Coelho from the Boeke lab, for assistance with WGS. This work is part of the Sc2.0 project (http://syntheticyeast.org/), supported by NSF grants MCB-1026068dslngs1, MCB-1443299, MCB-1616111, and MCB-1921641 to J.D.B. as well as the UKRI EPSRC Fellowship (EP/V033794/1) to G.S. J.D.B. S.L. and J.L. conceptualized the study. J.D.B. G.S. and J.S.B. supervised the study. S.L. J.L. L.L.-S. Y.Z. W.Z. M.A.B.H. L.H.M. and F.Y. performed the experiments, including synthetic DNA assembly, construction and analysis of intermediate synVIII strains, centromere transplantation, Hi-C library construction and data processing, and RNA sequencing and analysis. S.L.L. N.E. V.F. and E.L. performed whole-genome sequencing and analysis. Build-A-Genome class students contributed building block and minichunk DNA assembly. J.C. and J.S.B. curated the synVIII sequencing datasets. The manuscript was drafted by S.L. J.L. L.L.-S. and J.D.B. and edited by all authors. J.B. is a founder and director of CDI Labs, Inc. a founder of and consultant to Neochromosome, Inc. a founder, SAB member of, and consultant to ReOpen Diagnostics, LLC, and serves or served on the Scientific Advisory Board of the following: Logomix, Inc. Modern Meadow, Inc. Rome Therapeutics, Inc. Sample6, Inc. Sangamo, Inc. Tessera Therapeutics, Inc. and the Wyss Institute. J.S.B. is a founder of and consultant to Neochromosome. G.S. is a consultant to Neochromosome Inc. and ZenithAI. Publisher Copyright: {\textcopyright} 2023 The Authors",
year = "2023",
month = nov,
day = "8",
doi = "10.1016/j.xgen.2023.100437",
language = "English",
volume = "3",
journal = "Cell Genomics",
issn = "2666-979X",
publisher = "Cell Press",
number = "11",
}