title: Efficient Cellular Entry of (r-x-r)-Type Carbamate-Plasmid DNA Complexes and Its Implication for Noninvasive Topical DNA Delivery to Skin.
abstract: Arginine-rich cell penetrating peptides are powerful tools for in vitro as well as in vivo delivery of a wide plethora of biomolecules. However, presence of consecutive arginine residues leads to enhanced amenability for proteolytic degradation as well as steric hindrances for membrane interactions which compromise its bioavailability. In order to overcome these limitations we previously reported a safe and stable octaarginine based oligomer, i.e., (r-x-r)4-carbamate, where the backbone amide linkages were replaced by carbamate linkages and 6-aminohexanoic acid based spacer moieties were incorporated for better flexibility, hydrophobicity, optimal spacing of guanidinium groups, and protection against proteolytic cleavage; resulting in improved transfection efficiency over its amide counterpart. In the present work we have investigated the mechanism behind this enhanced transfection efficiency and, based on our observations, demonstrate how the synergistic effect of rationalized oligomer designing, complex characteristics, and cell type contributes to overall effective intracellular delivery. Our results indicate that the (r-x-r)4-carbamate-plasmid DNA complexes primarily utilize lipid raft dependent pathway of cellular entry more than other pathways, and this possibly facilitates their increased entry in the lipid raft rich milieu of skin cells. We also emphasize the utility of oligomer (r-x-r)4-carbamate as an efficient carrier for topical delivery of nucleic acids in skin tissue. This carrier can be utilized for safe, efficient, and noninvasive delivery of therapeutically relevant macromolecular hydrophilic cargo like nucleic acids to skin.
authors.affiliation: Department of Structural Biology, CSIR-Institute of Genomics and Integrative Biology , South Campus, Mathura Road, New Delhi, India 110020.|Academy of Scientific and Innovative Research (AcSIR) , Anusandhan Bhawan, 2 Rafi Marg, New Delhi, India 110001.
authors.forename: Manika
authors.initials: M
authors.lastname: Vij
authors.affiliation: Department of Structural Biology, CSIR-Institute of Genomics and Integrative Biology , South Campus, Mathura Road, New Delhi, India 110020.
authors.forename: Poornemaa
authors.initials: P
authors.lastname: Natarajan
authors.affiliation: CSIR-National Chemical Laboratory , Pune, Maharashtra, India 411008.
authors.forename: Amit K
authors.initials: AK
authors.lastname: Yadav
authors.affiliation: CSIR-National Chemical Laboratory , Pune, Maharashtra, India 411008.
authors.forename: Kiran M
authors.initials: KM
authors.lastname: Patil
authors.affiliation: Department of Structural Biology, CSIR-Institute of Genomics and Integrative Biology , South Campus, Mathura Road, New Delhi, India 110020.
authors.forename: Tanuja
authors.initials: T
authors.lastname: Pandey
authors.affiliation: Delhi Technological University , Shahbad Daulatpur, Main Bawana Road, New Delhi, India 110042.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Gupta
authors.affiliation: Delhi Technological University , Shahbad Daulatpur, Main Bawana Road, New Delhi, India 110042.
authors.forename: Deenan
authors.initials: D
authors.lastname: Santhiya
authors.affiliation: CSIR-National Chemical Laboratory , Pune, Maharashtra, India 411008.
authors.forename: Vaijayanti A
authors.initials: VA
authors.lastname: Kumar
authors.affiliation: CSIR-National Chemical Laboratory , Pune, Maharashtra, India 411008.
authors.forename: Moneesha
authors.initials: M
authors.lastname: Fernandes
authors.affiliation: Department of Structural Biology, CSIR-Institute of Genomics and Integrative Biology , South Campus, Mathura Road, New Delhi, India 110020.|Academy of Scientific and Innovative Research (AcSIR) , Anusandhan Bhawan, 2 Rafi Marg, New Delhi, India 110001.
authors.forename: Munia
authors.initials: M
authors.lastname: Ganguli
mesh_terms: D000818:Animals; D001120:Arginine / Q000378:metabolism; D016466:CHO Cells; D002219:Carbamates / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000378:metabolism; D003412:Cricetulus; D004247:DNA / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry; D009696:Nucleic Acids / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism*; D012867:Skin / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D002219:Carbamates; D057846:Cell-Penetrating Peptides; D008055:Lipids; D009696:Nucleic Acids; D009842:Oligopeptides; C448619:octaarginine; D004247:DNA; D001120:Arginine
keywords: cell penetrating peptides; endocytosis; nucleic acid delivery; skin penetration; transfection