研究人员介绍了一种高效形成单拷贝人类人工染色体（HAC）的方法。它采用了一个约750千碱基的构建体，该构建体足够大，可以容纳存在于内外着丝粒的不同染色质类型，从而避免了多聚化的需要。通过采用酵母原生质球融合技术，向哺乳动物细胞的输送得以简化。这些进展使得染色体工程化能够在后生动物细胞中忠实地进行。
据介绍，大型DNA组装方法是合成原核生物和酵母芽殖染色体的里程碑式成就的基础。芽殖酵母通过约125碱基对DNA序列定义的着丝粒控制染色体遗传，而哺乳动物和许多其他真核生物则使用大型表观遗传着丝粒。利用着丝粒表观遗传学可以形成HAC，但还不足以避免初始DNA分子在进入细胞后发生猖獗的多聚化。
附：英文原文
Title: Efficient formation of single-copy human artificial chromosomes
Author: Craig W. Gambogi, Gabriel J. Birchak, Elie Mer, David M. Brown, George Yankson, Kathryn Kixmoeller, Janardan N. Gavade, Josh L. Espinoza, Prakriti Kashyap, Chris L. Dupont, Glennis A. Logsdon, Patrick Heun, John I. Glass, Ben E. Black
Issue&Volume: 2024-03-22
Abstract: Large DNA assembly methodologies underlie milestone achievements in synthetic prokaryotic and budding yeast chromosomes. While budding yeast control chromosome inheritance through ~125-base pair DNA sequence-defined centromeres, mammals and many other eukaryotes use large, epigenetic centromeres. Harnessing centromere epigenetics permits human artificial chromosome (HAC) formation but is not sufficient to avoid rampant multimerization of the initial DNA molecule upon introduction to cells. We describe an approach that efficiently forms single-copy HACs. It employs a ~750-kilobase construct that is sufficiently large to house the distinct chromatin types present at the inner and outer centromere, obviating the need to multimerize. Delivery to mammalian cells is streamlined by employing yeast spheroplast fusion. These developments permit faithful chromosome engineering in the context of metazoan cells.
DOI: adj3566
Source: https://www.science.org/doi/10.1126/science.adj3566