Splice redirection as a convenient assay to monitor CPP-ON efficiency and mechanism.
Several strategies based on synthetic oligonucleotides (ON) have been proposed to control gene expression. As for most biomolecules, however, delivery has remained a major roadblock for in vivo applications. Conjugation of steric-block neutral DNA mimics, such as peptide nucleic acids (PNA) or phosphorodiamidate morpholino oligonucleotides (PMO), to cell-penetrating peptides (CPP) has recently been proposed as a new delivery strategy. It is particularly suitable for sequence-specific interference with pre-mRNA splicing, thus offering various applications in fundamental research and in therapeutics. The chemical synthesis of these CPP-ON conjugates will be described as well as easy-to-implement assays to monitor cellular uptake, endosome leakage, and efficiency of splicing redirection.
Université Montpellier, Montpellier, France.
Rachida
R
Abes
Andrey A
AA
Arzumanov
Amer F
AF
Saleh
Fatouma
F
Said Hassane
Michael J
MJ
Gait
Bernard
B
Lebleu
D002463:Cell Membrane Permeability; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry / Q000378:metabolism; D004705:Endocytosis; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008301:Maleimides / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020135:Peptide Nucleic Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics*; D012503:Saponins / Q000378:metabolism
D057846:Cell-Penetrating Peptides; D004220:Disulfides; D004337:Drug Carriers; D008081:Liposomes; D008301:Maleimides; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D020135:Peptide Nucleic Acids; D012503:Saponins; C043592:maleimide; D008156:Luciferases