Nanoscale structure of protamine/DNA complexes for gene delivery

Motta, S.; Brocca, Paola; Favero, Elena Del; Rondelli, Valeria; Cantù, Laura; Amici, Augusto; Pozzi, Daniela; Caracciolo, Giulio

doi:10.1063/1.4790588

Cited by 20 publications

(20 citation statements)

References 27 publications

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“…Protamines, responsible for sperm chromatin condensation, have been chosen for their precise amino acid composition and high charge density (21 over 30 amino acids are positively charged in salmon protamine). Soluble DNA-protamine complexes formed at low DNA and salt concentration as established in previous studies 26 27 28 remain stable for months. We focus here on the mechanisms involved in their formation and stability by using a combination of multiple experimental and simulation approaches.…”

supporting

confidence: 62%

A route to self-assemble suspended DNA nano-complexes

Lansac

Degrouard

Renouard

et al. 2016

Sci Rep

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Highly charged polyelectrolytes can self-assemble in presence of condensing agents such as multivalent cations, amphiphilic molecules or proteins of opposite charge. Aside precipitation, the formation of soluble micro- and nano-particles has been reported in multiple systems. However a precise control of experimental conditions needed to achieve the desired structures has been so far hampered by the extreme sensitivity of the samples to formulation pathways. Herein we combine experiments and molecular modelling to investigate the detailed microscopic dynamics and the structure of self-assembled hexagonal bundles made of short dsDNA fragments complexed with small basic proteins. We suggest that inhomogeneous mixing conditions are required to form and stabilize charged self-assembled nano-aggregates in large excess of DNA. Our results should help re-interpreting puzzling behaviors reported for a large class of strongly charged polyelectrolyte systems.

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supporting

confidence: 62%

A route to self-assemble suspended DNA nano-complexes

Lansac

Degrouard

Renouard

et al. 2016

Sci Rep

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“…This value for the Porod exponent reveals a typical feature of long-range clustering in biological macromolecules 41 and suggested structures with sharp interfaces and a surface fractal dimension of D s = 6 -3.6 = 2.4. 41,42 An intense peak at q 0 = 1.12 nm -1 , followed by its second order at q 1 = 2.25 nm -1 , indicated a repeat distance of d = 2/q 0 = 5.6 nm which can be ascribed to the separation between layers in a lamellar structure containing DNA in the interstices. Since the diameter of double-stranded DNA without a counter ion solvation shell was around 2 nm, 4 lamellae formed upon complexation can be smaller than rod-like micelles formed by the lipopeptide.…”

Section: Structural Analysis Of Peptide Complexation With Short Dna Fmentioning

confidence: 97%

Structural behaviour and gene delivery in complexes formed between DNA and arginine-containing peptide amphiphiles

et al. 2016

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We describe in depth the structure of complexes formed between DNA and two classes of arginine-containing peptide amphiphiles, namely, the lipopeptide PRW-C (P = proline, R = arginine, W = tryptophan, C = C16 : 0 alkyl chain) and the bolaamphiphile RFLFR (R = arginine, F = phenylalanine, L = leucine). A combination of X-ray and neutron scattering provided unprecedented insights into the local structure of these complexes. Lipopeptide-based complexes self-assembled into layered structures with large-scale fractal features, hosting DNA in the interstices. Bola-amphiphile scaffolds were characterized by planar structures with DNA strands presumably sandwiched in-between peptide nanotapes. Importantly, complexation did not affect the structural integrity of DNA in either of the two complexes. The bolaamphiphile conjugates displayed high levels of molecular ordering in contrast to the liquid-crystalline features observed in lipopeptide assemblies. Peptide-DNA complexes were assessed for their potential as a means to deliver the reporter vector pEGFP-N1 into SW480 human colon carcinoma cells. Successfully transfected cells expressed green fluorescent protein. The potentiating effect of PRW-C on the cellular uptake of ectopic DNA was found to be much greater than that observed with RFLFR. In contrast to the bolaamphiphile-based conjugate, the liquid-crystalline nature of the lipopeptide complex is likely to play a key role in DNA release and transfection efficiency since these weakly bound structures require lower energy expenditure during disassembly and load release.

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“…limits the complex to cross the cellular membrane (11). Therefore, there is absolutely essential to facilitate an electrostatic interaction between protamine and the negative charge on the cell membrane.…”

Section: Discussionmentioning

confidence: 99%

“…In order to promote the transfection process, it is desired to facilitate the electrostatic interaction of the transfection reagent with the negative charge of DNA backbone and the cell membrane (11). Also, previous data has been shown that precondensation of DNA by an additional cationic charge improves its protection against degradation (12).…”

Section: Introductionmentioning

confidence: 99%

High-efficiency Non-viral Transfection of Human Primary T Lymphocytes

Khanahmad¹,

Rahimmanesh²,

Totonchi³

2020

Preprint

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The development and optimization of an effective non-viral gene delivery method for genetic manipulation of primary human T cells is a major challenge in clinical immunotherapy researches. According to the low transfection efficiency of conventional methods in human primary T cells, there is an effort in order to increase the transfection rate in these cells. Protamine is an FDA-approved compound with a documented safety profile that enhances DNA condensation for gene delivery. In this study, the effect of protamine on the transfection efficiency of standard transfection reagents, TurboFect, and Lipofectamine 2000 was evaluated in order to transfect primary human T cells. Results demonstrated that protamine condenses DNA and increases the positive charge of DNA/Cargo complex efficiently without any cytotoxic effect on the primary human T cells. The results also revealed that the DNA/Protamine/Cargo complexes effectively transfect human primary T cells.

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Nanoscale structure of protamine/DNA complexes for gene delivery

Cited by 20 publications

References 27 publications

A route to self-assemble suspended DNA nano-complexes

A route to self-assemble suspended DNA nano-complexes

Structural behaviour and gene delivery in complexes formed between DNA and arginine-containing peptide amphiphiles

High-efficiency Non-viral Transfection of Human Primary T Lymphocytes

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