New generation of efficient peptide-based vectors, NickFects, for the delivery of nucleic acids.
Harnessing of a branched structure is a novel approach in the design of cell-penetrating peptides and it has provided highly efficient transfection reagents for intracellular delivery of nucleic acids. The new stearylated TP10 analogs, NickFects, condense plasmid DNA, splice correcting oligonucleotides and short interfering RNAs into stable nanoparticles with a size of 62-160nm. Such nanoparticles have a negative surface charge (-11 to -18mV) in serum containing medium and enable highly efficient gene expression, splice correction and gene silencing. One of the novel peptides, NickFect51 is capable of transfecting plasmid DNA into a large variety of cell lines, including refractory suspension and primary cells and in several cases exceeds the transfection level of commercially available reagent Lipofectamine™ 2000 without any cytotoxic side effects. Additionally we demonstrate the advantages of NickFect51 in a protein production system, QMCF technology, for expression and production of recombinant proteins in hardly transfectable suspension cells.
Institute of Technology, University of Tartu, Tartu, Estonia. piret.arukuusk@ut.ee
Piret
P
Arukuusk
Ly
L
Pärnaste
Nikita
N
Oskolkov
Dana-Maria
DM
Copolovici
Helerin
H
Margus
Kärt
K
Padari
Kaidi
K
Möll
Julia
J
Maslovskaja
Radi
R
Tegova
Gaily
G
Kivi
Andres
A
Tover
Margus
M
Pooga
Mart
M
Ustav
Ulo
U
Langel
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D009696:Nucleic Acids / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D013229:Stearic Acids / Q000737:chemistry; D014162:Transfection / Q000379:methods
D057846:Cell-Penetrating Peptides; D009696:Nucleic Acids; D009841:Oligonucleotides; D034741:RNA, Small Interfering; D013229:Stearic Acids; C031183:stearic acid; D008156:Luciferases