Abstract
Four cationic chiral amino acid-based surfactants, cis- and trans-1 and cis- and trans-2, have been studied as DNA-condensing agents with enhanced properties and the absence of cell toxicity. The polar head of the surfactant is made of a cyclobutane β-amino acid in which the amino group is a hydrochloride salt and the carboxyl group is involved in an amide bond, allowing the link with hydrophobic C12 (surfactant 1) or C16 (surfactant 2) chains. The ability of these surfactants to condense DNA was investigated using a dye exclusion assay, gel electrophoresis, and circular dichroism and compared with the well-studied dodecyltrimethylammonium bromide (DTAB) and cetyltrimethylammonium bromide (CTAB). The surfactant with the longest chain length and the trans stereochemistry (trans-2) was found to be the most efficient in condensing the DNA, including CTAB. Surfactant cis-2 was found to be less efficient, probably due to its poorer solubility. The β-amino acid surfactants with the shorter chain length behaved similarly, such that the cis/trans stereochemistry does not seem to play a role in this case. Interestingly, these were also found to induce DNA condensation for the same concentration as trans-2 and CTAB but showed a lower binding cooperativity. Therefore, a longer alkyl chain only slightly improved the effectiveness of these surfactants. Further, atomic force microscopy revealed that they compact DNA into small complexes of about 55-110 nm in diameter.
Highlights
Gene therapy has gained significant attention over the past two decades as a potential method for treating genetic disorders such as Severe Combined Immunodeficiency,[1] Hemophilia B,2 cystic fibrosis,[3] Leber’s Congenital Amaurosis,[4] and Parkinson’s disease[5] as well as an alternative method to traditional chemotherapy for treating cancer.[6]
The results obtained for the novel cis- and trans-1 and -2 surfactants were compared with those of commercial dodecyltrimethylammonium bromide (DTAB) and cetyltrimethylammonium bromide (CTAB), which have been extensively studied as DNA-compacting agents
The obtained results were complemented with Electrophoretic Mobility Shift Assay (EMSA) experiments that explore the degree of complexation of DNA by the cationic surfactants, while circular dichroism (CD) spectroscopy was used to monitor the variation of the DNA structure upon surfactant self-assembly
Summary
Gene therapy has gained significant attention over the past two decades as a potential method for treating genetic disorders such as Severe Combined Immunodeficiency,[1] Hemophilia B,2 cystic fibrosis,[3] Leber’s Congenital Amaurosis,[4] and Parkinson’s disease[5] as well as an alternative method to traditional chemotherapy for treating cancer.[6] It consists of introducing a target gene, which is encoded in a DNA or RNA chain, into a specific cell. Fundamental problems associated with viral vector systems, including toxicity, immunogenicity, and limitations with respect to scaled-up procedures, encouraged the investigation of other potential vectors for introducing the DNA into the targeted tissues.[9,10] The role of chemists in the field of gene therapy is to design and prepare new nonviral vectors, based on cationic lipids,[11−13] cationic surfactants,[12,14] cationic polymers,[15,16] metal cations,[17] dendrimers,[18,19] polypeptides,[20,21] and nanoparticles.[22,23]
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