Nucleic acid-matrix attachment recognition regions--as facilitators in plasmid transfer.
Non-viral gene transfer is an alternative to viral vectors for gene transfer. However, non-viral transgene expression remains undesirably low and transient. Matrix attachment regions (MARs) are DNA elements that are defined by their high affinity for the nuclear matrix. MARs may also be related to long-term transgene expression in vitro. The purpose of this research is to evaluate human interferon-beta MARs element in various cell types. This was accomplished by constructing MARs-containing plasmid DNA (pDNA) and comparing their transgene expression with non-MARs-containing pDNA. We found that MARs-containing pDNA increased and prolonged the expression in Chinese hamster ovary (CHO) cells, but not in human neuroblastoma cells (SKnSH) and neuronal cells (primary neuron, astroglia and microglia). From the cotransfection experiment, MARs-containing pDNA had a trans effect on another pDNA expression. A PCR method was used to monitor the intracellular distribution of pDNA after cellular fractionalization. We found that non-MAR containing pDNA demonstrates similar intracellular distribution as non-MARs-containing pDNA.
Department of Pharmaceutics, College of Pharmacy, University of Florida, Gainesville 32603, USA.
Pattravadee
P
Chancham
Tessa
T
van Ijperen
Issam
I
McDoom
Jeffrey A
JA
Hughes
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D016466:CHO Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D006224:Cricetinae; D006801:Humans; D045170:Matrix Attachment Regions / Q000235:genetics*; D009696:Nucleic Acids / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D014162:Transfection / Q000379:methods*
D009696:Nucleic Acids