Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations.

Felgner, Jiin; Kumar, Raj; Sridhar, C.; Wheeler, Carl J.; Tsai, Ya-Li; Border, R.; Ramsey, Patrick S.; Martin, Michael U.; Felgner, Philip L.

doi:10.1016/s0021-9258(17)41980-6

Cited by 1,198 publications

(363 citation statements)

References 42 publications

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“…We were prompted to pursue this question because of an electron microscopy study done by our group showing a larger fraction of DNA-free liposomes when SUV are used to complex DNA instead of sedimented multilamellar vesicles (sMLV) (Jang and Heath, 1997). Additional motivation for this study came from the lack of characterization studies using MLV lipoplexes, and from the well established knowledge that these are better transfecting agents than SUV lipoplexes (Felgner et al, 1994;Liu et al, 1997;Ross et al, 1998;MacDonald et al, 1999;Ross and Hui, 1999;Zuidam et al, 1999), thus making their characterization an essential step toward the development of more efficient gene delivery systems.…”

Section: Introductionmentioning

confidence: 99%

The Effect of Liposome Size on the Final Lipid/DNA Ratio of Cationic Lipoplexes

Gonçalves

Debs

Heath

2004

Biophysical Journal

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Several studies have demonstrated that lipoplexes are two-phase systems over most mixing lipid/DNA charge ratios. Because these studies have focused on small unilamellar vesicles (SUV), they leave open the question as to whether a similar pattern is followed by other liposome types. The main purpose of this work is to examine the question further by characterizing the assembly of cationic lipoplexes prepared from 1-[2-(oleoyloxy)ethyl]-2-oleyl-3-(2-hydroxyethyl)imidazolinium chloride (DOTIM)/dioleoylphosphatidylethanolamine (DOPE) (1:1) liposomes of various types. Sedimentation in sucrose density gradients reveals that large unilamellar vesicles (LUV) and sedimented multilamellar vesicles (sMLV), as opposed to SUV, form lipoplexes that exist as a single phase over a relatively broad range of mixing (+/-) ratios. This is indicated by observing that most of the LUV and sMLV become involved in the assembly reaction up to mixing (+/-) ratios of 4 and 9, respectively, while only a small and constant fraction of SUV associates with DNA at all mixing (+/-) ratios tested. Consequently, while maximal (+/-) ratios of approximately 4.5 and 9 are found in LUV and sMLV lipoplexes, respectively, a final (+/-) ratio of only approximately 2 is determined in SUV lipoplexes. Isothermal titration calorimetry shows that this is the lowest possible charge ratio achieved when liposomes are titrated with DNA. Based on these observations and on the size differences of the liposomes used, a model of lipoplex formation is proposed.

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Section: Introductionmentioning

confidence: 99%

The Effect of Liposome Size on the Final Lipid/DNA Ratio of Cationic Lipoplexes

Gonçalves

Debs

Heath

2004

Biophysical Journal

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“…This was a nucleocapsid based nucleic acid vaccine that induced cross-subtype protection against both group 1 and group 2 viruses (A/PR/8/34 (H1N1) and A/HK/68 (H3N2)). The utility of cationic lipids for gene delivery was discovered and reported in 1987 [17] and synthetic self-assembling lipoplexes for gene delivery described [18][19][20]. These results spawned a branch of gene therapy science, and an NIH study section, Genes and Drug Delivery (GDD) was established in 2002 that continues to support this research emphasis.…”

Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 virus masking of RBD epitopes; unmasking and cross-reactivity induced by mRNA vaccines

Assis

Jain

Nakajima

et al. 2021

Preprint

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We analyzed data from two ongoing COVID-19 longitudinal serological surveys in Orange County, CA., between April 2020 and March 2021. A total of 8,476 finger stick blood specimens were collected before and after an aggressive mRNA vaccination campaign. IgG levels were determined using a multiplex antigen microarray containing 10 SARS-CoV-2 antigens, 4 SARS, 3 MERS, 12 Common CoV, and 8 Influenza antigens. Twenty-six percent of 3,347 specimens from unvaccinated Orange County residents in December 2020 were SARS-CoV-2 seropositive. The Ab response was predominantly against nucleocapsid (NP), full length spike and the spike S2 domain. Anti-receptor binding domain (RBD) reactivity was low and there was no cross-reactivity against SARS S1 or SARS RBD. An aggressive mRNA vaccination campaign at the UCI Medical Center started on December 16, 2020 and 6,724 healthcare workers were vaccinated within 3 weeks. Seroprevalence increased from 13% in December to 79% in January, 93% in February and 99% in March. mRNA vaccination induced much higher Ab levels especially against the RBD domain and significant cross-reactivity against SARS RBD and S1 was also observed. Nucleocapsid protein Abs can be used to distinguish individuals in a population of vaccinees to classify those who have been previously infected and those who have not, because nucleocapsid is not in the vaccine. Previously infected individuals developed higher Ab titers to the vaccine than those who have not been previously exposed. These results indicate that mRNA vaccination rapidly induces a much stronger and broader Ab response than SARS-CoV-2 infection.

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“…The evidence illustrated the possibility to elevate the stability of mRNA for the purpose of producing more proteins (Hayashi, Lamba, Slowik, Reh, & Bermingham‐McDonogh, 2010; Mockey et al, 2006; F. Zohra et al, 2007). Thus, the translation products of mRNAs were enhanced by incorporating sequences such as β‐globin and Venezuelan equine encephalitis virus to stabilize the mRNA or by improving transfection reagents (Felgner et al, 1994; Hayashi et al, 2010; Holtkamp et al, 2006; Kariko, Kuo, & Barnathan, 1999).…”

Section: Discussionmentioning

confidence: 99%

Liposome‐mediated messenger RNA: An alternative for fish cell transfection in culture

et al. 2020

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Liposome‐mediated plasmid DNA (pDNA) transfection is a relatively efficient transfection method that depends highly on actively dividing cells. However, numerous fish cells are mitotically inactive under in vitro culture conditions, thus limiting the application of liposome‐mediated pDNA transfection. Liposome‐mediated mRNA transfection is an attractive alternative in post‐mitotic cells because it eliminates the transfer of genes into the nucleus and therefore is independent of cell proliferation. In this study, pDNA is compared with mRNA encoding biomarker proteins (enhanced green fluorescent protein [eGFP] and luciferase) mediated by cationic liposome in zebrafish embryo fibroblast cell line (ZF4), with or without proliferation restriction, and in mitotically inactive turbot liver primary cells. The results indicated that mRNA encoding eGFP exerted much higher transfection efficiency than pDNA in ZF4 cells. Moreover, mRNA encoding eGFP was detected and disappeared sooner than pDNA. Chemiluminescence of the biomarker protein luciferase showed that liposome‐mediated mRNA transfection translated about threefold more protein without proliferation restriction and about tenfold more protein with proliferation restriction than pDNA transfection. In turbot liver cells, mRNA induced higher transfection efficiency and greater luciferase protein expression than pDNA. Greater PPARα expression and expression of downstream target genes were induced by mRNA rather than by pDNA transfection. In conclusion, cationic liposome‐mediated mRNA is an alternative for fish cell transfection due to higher transfection efficiency and protein expression levels and faster translation onset.

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Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations.

Cited by 1,198 publications

References 42 publications

The Effect of Liposome Size on the Final Lipid/DNA Ratio of Cationic Lipoplexes

The Effect of Liposome Size on the Final Lipid/DNA Ratio of Cationic Lipoplexes

SARS-CoV-2 virus masking of RBD epitopes; unmasking and cross-reactivity induced by mRNA vaccines

Liposome‐mediated messenger RNA: An alternative for fish cell transfection in culture

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