Efficient formation of single-copy human artificial chromosomes.
Science
; 383(6689): 1344-1349, 2024 Mar 22.
Article
em En
| MEDLINE
| ID: mdl-38513017
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Centrômero
/
Cromossomos Artificiais Humanos
/
Epigênese Genética
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article