Lentiviral transduction of neonatal rat ventricular myocytes preserves ultrastructural features of genetically modified cells

Virology. 2021 Oct:562:190-196. doi: 10.1016/j.virol.2021.07.015. Epub 2021 Jul 31.

Abstract

Preserving morphological features that are important for cell function and structure is a critical parameter for in vitro experiments with rat cardiomyocytes. Lentiviral vectors are commonly used as gene transfer tool because of its high flexibility, efficiency to deliver expression cassettes and versatility of transducing quiescent cells. The tropism of the recombinant viral particle can be determined depending on the virus envelope, which shows a specific binding to cell surface receptors on the target cell. The combination of promoter arrangement and viral envelope must be optimized to achieve a greater transduction efficiency and a higher transgene expression. In this study we explored the optimization of promoters and heterologous envelopes to transduce primary culture of neonatal rat ventricular myocytes. Our results suggest a robust expression driven by the cytomegalovirus promoter, and high efficiency transduction mediated by VSV-G envelope with no apparent compromising ultrastructural features of genetically modified cells.

Keywords: Cardiomyocyte; Gene transfer; Lentivirus; Pseudotyping; Superinfection interference.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cells, Cultured
  • Cytomegalovirus / genetics
  • Gene Expression
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lentivirus / genetics*
  • Membrane Glycoproteins / genetics
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism
  • Promoter Regions, Genetic
  • Rats
  • Sarcomeres / ultrastructure
  • Transduction, Genetic / methods*
  • Transgenes
  • Viral Envelope Proteins / genetics
  • Viral Pseudotyping

Substances

  • G protein, vesicular stomatitis virus
  • Membrane Glycoproteins
  • Viral Envelope Proteins
  • Green Fluorescent Proteins