Constructing Nucleic Acid Delivering Lipoproteoplexes from Coiled-Coil Supercharged Protein and Cationic Liposomes

Joseph Thomas, Jonathan Sun, Jin Kim Montclare

Research output: Chapter in Book/Report/Conference proceedingChapter


The safe and efficient delivery of nucleic acids is crucial for both clinical applications of gene therapy and pre-clinical laboratory research. Such delivery strategies rely on vectors to condense nucleic acid payloads and escort them into the cell without being degraded in the extracellular environment; however, the construction and utilization of these vectors can be difficult and time-consuming. Here, we detail the steps involved in the rapid, laboratory-scale production and assessment of a versatile, nucleic acid delivery vehicle, known as the lipoproteoplex. In this chapter, we outline: (1) the recombinant synthesis and subsequent purification of the supercharged coiled-coil protein component known as N8; (2) the synthesis of cationic liposomes from dioleoyl-3-trimethylammonium propane (DOTAP) and sodium cholate; (3) and finally a protocol for the delivery of a model siRNA cargo into a cultured cell line.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages17
StatePublished - 2024

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Gene delivery
  • Lipoproteoplex
  • Liposome
  • Protein engineering
  • Recombinant expression
  • Transfection

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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