Context-dependent neocentromere activity in synthetic yeast chromosome VIII
Stephanie Lauer, Jingchuan Luo, Luciana Lazar‐Stefanita, Weimin Zhang, Laura H. McCulloch, Viola Fanfani, Evgenii Lobzaev, Max A. B. Haase, Nicole Easo, Yu Zhao, Fangzhou Yu, Jitong Cai, Lajari Anne, James S. Barger, Naz Belkaya, Kristin Boulier, Kirk Butler, Melanie M. Callaghan, Calvin Chang, Janice Chen, Xueni Jennifer Chen, In Young Cho, Elliot H. Choi, Woo Jin Choi, James Chuang, Ashley L. Cook, Eric M. Cooper, Nicholas Timothy Culbertson, Jessilyn Dunn, Charlotte E. Floria, Breeana Grogan Anderson, Nathalie P. Held, Emily Hsiao, Joseph Igwe, K. Kang, Joana Karanxha, Marie Kelly, Arjun Khakhar, Chachrit Khunsriraksakul, John J. Kim, Dong Won Kim, Jin Wan Kim, Alex Lamb, David Sung Han Lee, Yoon Kyung Lee, Jongseuk Lim, Steffi Liu, Jeremy López, Zhen A. Lu, Henry Ma, Jordan A. Mandel, Jessica Mao, Jordan Matelsky, Jonathan Merran, Rishikesh Mohan, Christopher Montoya, Sindurathy Murugan, Lisa Ni, Won Chan Oh, Youngrok Park, Laura C. Paulsen, Nick A. Phillips, Sudarshan Pinglay, Vikram Aditya Rajan, Garrett Ransom, Erin Rhoads, Praneeth Sanna, Emily Scher, Jinesh Shah, Ashwyn K. Sharma, Maya C. Shepardson, Joanne Song, Sainikhil Sontha, Venkatesh Srinivas, Scott Tan, Ang A. Tu, Skyler Uhl, Wang Xiaoyue, Fangzhou Yu, Justine R. Yu, Amadeus Zhu, Joel S. Bader, Giovanni Stracquadanio, Jef D. Boeke
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.