Baculovirus-Mediated Gene Expression in Chicken Primary Cells

Abstract

A recombinant baculovirus was constructed containing an expression cassette with a reporter gene, green fluorescent protein, directed by a constitutive mammalian promoter: a human cytomegalovirus immediate early promoter/enhancer (CMV-IE). High titer virus was prepared with ultracentrifugation. Efficient gene delivery and expression were observed in the virus-treated chicken primary culture, myoblast cells, and whole embryonic fibroblast cells. It was noticed that an addition of sodium butyrate (a selective histone deacetylase inhibitor) to viral transduction medium extremely enhanced the reporter-gene expression. However, there is no effect of presence of trichostatin A observed. To maximize the reporter-gene expression, the baculoviral infection condition was optimized with both cell types. Our approaches demonstrated that recombinant baculovirus could efficiently deliver its genome DNA into chicken primary cells and that CMV-IE, a mammalian-cell-active promoter, was functional in chicken primary cells and could direct a high level of gene expression. Clearly, the recombinant baculovirus provides an alternative means for foreign gene delivery into avian cells.

Serum-free transfection of CHO cells with chemically defined transfection systems and investigation of their potential for transient and stable transfection

“…In general, improvements in transient gene expression are multiplex with the most prominent to be the transfection efficiency, the maximum cell density and the concentration of protein enhancers (Jardin et al 2008). Other enhancers are sodium butyrate (Leisy et al 2003;Mahonen et al 2007;Ping et al 2006) and trichostatin A (Spenger et al 2004). More recently proto-oncogenes, cell cycle control genes, growth factor genes and anti-apoptotic genes have been used to improve production hosts or transient expression (Wurm 2004;Backliwal et al 2008).…”

Improving heterologous protein expression in transfected Drosophila S2 cells as assessed by EGFP expression

“…Based on published studies (Dorner et al 1989;Chen et al 2002;Li and Li 2006;Zhao et al 2006;Ping et al 2006;Song et al 2006), sodium butyrate (NaBu), is a histone deacetylase inhibitor, affecting the chromatin structure and leading to the destabilization of nucleosomal structure. As a consequence, the NaBU influence on chromatin exposure may facilitate binding of transcription factors to DNA resulting in an increased and deregulated cell protein synthesis.…”

“…In this context, the transfection procedure, aiming the introduction of foreign DNA into cells, is an important step. The calcium phosphate method has been recommended for gene transfection of S2 cells (Van der Straten et al 1989) but procedures based on liposome reagent have been reported to be more efficient (Han 1996;Park et al 1999;Shin and Cha 2002) Besides the metallothionein promoter induction by copper sulfate (CuSO 4 ) (Huang et al 2004) it has been studied the enhancement of gene expression by agents, such as the sodium butyrate (NaBu), a histone deacetylase inhibitor that lead to chromatin exposure and allow the transcriptional factors binding and increased gene expression (Dorner et al 1989;Li and Li 2006;Zhao et al 2006;Ping et al 2006;Song et al 2006).…”

Transgene expression in Penaeus monodon cells: evaluation of recombinant baculoviral vectors with shrimp specific hybrid promoters

“…After a long history, in 1985, first successful in vitro gene transfer by a recombinant baculovirus was accomplished by Carbonell et al (1985) that made baculovirus a tool in gene transfer technology, especially in the over expression of cloned genes (O'Reilly et al 1992). Since then, the recombinant baculoviruses have been successfully used for gene transfer in various eukaryotic cells including those of fishes (Smith and Summers 1989;Leisy et al 2003;Wagle and Jesuthasan 2003;Wagle et al 2004), chicken and duck (Ping et al 2006;Song et al 2006), fruit fly (Oppenheimer et al 1999; Lee et al 2000), honey bees (Ando et al 2007), shrimp (Lu et al 2005;Syed Musthaq et al 2009; Syed Musthaq and Kwang 2011) rabbit (Airenne et al 2000), monkey (Tani et al 2001) and human (Hofmann et al 1995;Kost and Condreay 2002). Besides, recombinant baculovirus has been used for gene therapy (Luo et al 2011;Zhao et al 2012), and vaccine production (van Oers 2006;Treanor et al 2007;Hu et al 2008;Cox 2012).…”