Rapid assembly of functional modules for generating human artificial chromosome constructs compatible with epigenetic centromere seeding.

ABSTRACT: The ongoing development of human artificial chromosomes (HACs) will permit investigation into essential centromere processes and the means to deliver large genetic cargoes to target cells. Starting with large (~750 kb) yeast artificial chromosome (YAC)-based constructs limits the rampant multimerization that has complicated many prior types of HACs. Large YAC construction is accomplished using transformation-associated recombination (TAR) strategies that can become unwieldly when several functional modules are to be incorporated and tested. To address this issue, we developed an approach where modules are built using high-fidelity in vitro assembly strategies in a bacterial artificial chromosome (BAC) format. Then, the assembled modules are transferred in a simplified TAR step into a recip

SUBMITTER: Birchak GJ 

PROVIDER: S-EPMC12669295 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature