Reprogramming a GFP reporter gene subjects it to complex lentiviral gene regulation

Methods Mol Biol. 2012;813:85-106. doi: 10.1007/978-1-61779-412-4_5.

Abstract

Late human immunodeficiency virus (HIV)-derived RNAs encoding relevant therapeutic targets or promising vaccine compounds, such as the HIV-1 group-specific antigen (Gag), are translocated from the nucleus into the cytoplasm via sophisticated export machinery. Relevant steps include the concerted action of several cis-acting RNA elements with the viral Rev-shuttle protein and several cellular components (Ran1/Exportin; Crm1). Based on detailed understanding of the molecular mechanisms guiding this complex process, we used rational codon usage modification to design and reprogram a GFP encoding reporter RNA now exactly mimicking the complex transcriptional and posttranscriptional regulation of late lentiviral mRNAs.

MeSH terms

  • Active Transport, Cell Nucleus
  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • DNA / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Gene Expression Regulation, Viral / genetics*
  • Genes, Reporter / genetics*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics*
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Lentivirus / genetics*
  • Molecular Sequence Data
  • Plasmids / genetics
  • Synthetic Biology / methods*

Substances

  • Green Fluorescent Proteins
  • DNA