Re-engineering an alphoid(tetO)-HAC-based vector to enable high-throughput analyses of gene function.

ABSTRACT: Human artificial chromosome (HAC)-based vectors represent an alternative technology for gene delivery and expression with a potential to overcome the problems caused by the use of viral-based vectors. The recently developed alphoid(tetO)-HAC has an advantage over other HAC vectors because it can be easily eliminated from cells by inactivation of the HAC kinetochore via binding of tTS chromatin modifiers to its centromeric tetO sequences. This provides unique control for phenotypes induced by genes loaded into the alphoid(tetO)-HAC. However, inactivation of the HAC kinetochore requires transfection of cells by a retrovirus vector, a step that is potentially mutagenic. Here, we describe an approach to re-engineering the alphoid(tetO)-HAC that allows verification of phenotypic changes attribu

SUBMITTER: Kononenko AV 

PROVIDER: S-EPMC3664798 | biostudies-literature | 2013 May

REPOSITORIES: biostudies-literature