Impacts of Nucleosome Positioning Elements and Pre-Assembled Chromatin States on Expression and Retention of Transgenes.

Autor:	Kwizera R; Department of Biochemistry, Purdue University, West Lafayette, IN 47907, USA., Xie J; Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47907, USA., Nurse N; Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47907, USA., Yuan C; Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN 47907, USA., Kirchmaier AL; Department of Biochemistry, Purdue University, West Lafayette, IN 47907, USA.
Jazyk:	angličtina
Zdroj:	Genes [Genes (Basel)] 2024 Sep 21; Vol. 15 (9). Date of Electronic Publication: 2024 Sep 21.
DOI:	10.3390/genes15091232
Abstrakt:	Background/objectives: Transgene applications, ranging from gene therapy to the development of stable cell lines and organisms, rely on maintaining the expression of transgenes. To date, the use of plasmid-based transgenes has been limited by the loss of their expression shortly after their delivery into the target cells. The short-lived expression of plasmid-based transgenes has been largely attributed to host-cell-mediated degradation and/or silencing of transgenes. The development of chromatin-based strategies for gene delivery has the potential to facilitate defining the requirements for establishing epigenetic states and to enhance transgene expression for numerous applications. Methods: To assess the impact of "priming" plasmid-based transgenes to adopt accessible chromatin states to promote gene expression, nucleosome positioning elements were introduced at promoters of transgenes, and vectors were pre-assembled into nucleosomes containing unmodified histones or mutants mimicking constitutively acetylated states at residues 9 and 14 of histone H3 or residue 16 of histone H4 prior to their introduction into cells, then the transgene expression was monitored over time. Results: DNA sequences capable of positioning nucleosomes could positively impact the expression of adjacent transgenes in a distance-dependent manner in the absence of their pre-assembly into chromatin. Intriguingly, the pre-assembly of plasmids into chromatin facilitated the prolonged expression of transgenes relative to plasmids that were not pre-packaged into chromatin. Interactions between pre-assembled chromatin states and nucleosome positioning-derived effects on expression were also assessed and, generally, nucleosome positioning played the predominant role in influencing gene expression relative to priming with hyperacetylated chromatin states. Conclusions: Strategies incorporating nucleosome positioning elements and the pre-assembly of plasmids into chromatin prior to nuclear delivery can modulate the expression of plasmid-based transgenes.
Databáze:	MEDLINE
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