Chapter 20
Lipofection-Based Delivery of DNA Vaccines
Monika Rak, Anna Go ´ ra-Sochacka, and Zbigniew Madeja
Abstract
The preventive and therapeutic potential of DNA vaccines combined with benefits of lipid-based delivery
(lipofection) allow efficient nucleic acid transfer and immunization applicable in treatment of infections,
cancer or autoimmune disorders. Lipofecting compositions consisting of cationic and neutral lipids can be
used for both in vitro and in vivo applications and may also play the role of adjuvants. Here we describe a
simple protocol of DNA vaccine carrier preparation based on cationic polyprenyl derivatives (PTAI—
trimethylpolyprenylammonium iodides) and commonly used helper lipids with use of basic laboratory
equipment. Such formulas have proven effective for immunization of animals as well as for cell transfection.
Key words Nonviral gene delivery, Lipofection, Lipid-based vaccine carriers, Polyprenyl-based vaccines, PTAI
1 Introduction
Wide preventive and therapeutic potential of DNA vaccines
includes viral, bacterial, and parasitic diseases as well as cancer,
autoimmune diseases and allergies. DNA biocompatibility, costefficient production, and good long-term storage properties are
their most important advantages. Moreover, nucleic acids may
also serve as vaccine adjuvants [1]. However, safety issues [2] and
poor immunogenicity in humans [3] are major concerns. Lipidbased solutions have been employed to address these drawbacks for
years [4–8] and still continue to be useful not only in DNA [9] but
also in RNA [10] settings.
Lipofection can be easily modified to meet case-specific
requirements via lipid composition and preparation [11, 12]. Lipofection strategy is based on cationic lipids that ensure spontaneous
interactions with negatively charged nucleic acids to form complexes called lipoplexes (see Fig. 1). Additional cationic and neutral
lipids (also designated as helper or co-lipids) modulate properties of
lipoplexes. DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolaBlaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_20, © Springer Science+Business Media, LLC, part of Springer Nature 2021
391
Lipofection-Based Delivery of DNA Vaccines
Monika Rak, Anna Go ´ ra-Sochacka, and Zbigniew Madeja
Abstract
The preventive and therapeutic potential of DNA vaccines combined with benefits of lipid-based delivery
(lipofection) allow efficient nucleic acid transfer and immunization applicable in treatment of infections,
cancer or autoimmune disorders. Lipofecting compositions consisting of cationic and neutral lipids can be
used for both in vitro and in vivo applications and may also play the role of adjuvants. Here we describe a
simple protocol of DNA vaccine carrier preparation based on cationic polyprenyl derivatives (PTAI—
trimethylpolyprenylammonium iodides) and commonly used helper lipids with use of basic laboratory
equipment. Such formulas have proven effective for immunization of animals as well as for cell transfection.
Key words Nonviral gene delivery, Lipofection, Lipid-based vaccine carriers, Polyprenyl-based vaccines, PTAI
1 Introduction
Wide preventive and therapeutic potential of DNA vaccines
includes viral, bacterial, and parasitic diseases as well as cancer,
autoimmune diseases and allergies. DNA biocompatibility, costefficient production, and good long-term storage properties are
their most important advantages. Moreover, nucleic acids may
also serve as vaccine adjuvants [1]. However, safety issues [2] and
poor immunogenicity in humans [3] are major concerns. Lipidbased solutions have been employed to address these drawbacks for
years [4–8] and still continue to be useful not only in DNA [9] but
also in RNA [10] settings.
Lipofection can be easily modified to meet case-specific
requirements via lipid composition and preparation [11, 12]. Lipofection strategy is based on cationic lipids that ensure spontaneous
interactions with negatively charged nucleic acids to form complexes called lipoplexes (see Fig. 1). Additional cationic and neutral
lipids (also designated as helper or co-lipids) modulate properties of
lipoplexes. DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolaBlaine A. Pfeifer and Andrew Hill (eds.), Vaccine Delivery Technology: Methods and Protocols, Methods in Molecular Biology,
vol. 2183, https://doi.org/10.1007/978-1-0716-0795-4_20, © Springer Science+Business Media, LLC, part of Springer Nature 2021
391