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Oxford대학교의 Wood 그룹과 Gait 그룹에서 2008년 11월에 파나진의 PNA를 이용한 연구 논문을 Nucleic Acids Research에 발표했습니다.
치료용으로 흔히 쓰이는 CPP인 (R-Ahx-R)4보다 (Ahx(X) = aminohexanoyl) PNA-internalization peptides (Pip)인 Pip2b와 접합한 PNA를 쓰면 같은 PNA농도에서도 3~4배 효과가 더 좋다고 보고했습니다. Pip2b-PNA가 세포 안의 endosomal trapping에서 더 잘 빠져나오기 때문에 효과가 좋은 것 같다고 발표하였습니다.
자세한 내용은 다음을 참고하세요.
Nucleic Acids Research 2008 36(20):6418-6428; doi:10.1093/nar/gkn671
http://nar.oxfordjournals.org/cgi/content/abstract/36/20/6418
Improved cell-penetrating peptide–PNA conjugates for splicing redirection
in HeLa cells and exon skipping in mdx mouse muscleGabriela D. Ivanova1, Andrey Arzumanov1, Rachida Abes2, Haifang Yin3, Matthew J. A. Wood3, Bernard Lebleu2 and Michael J. Gait1,*
1Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH UK, 2UMR 5235 CNRS, Université Montpellier 2, Place Eugene Bataillon, 34095 Montpellier cedex 5, France and 3Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK
Abstract
Steric blocking peptide nucleic acid (PNA) oligonucleotides have been used increasingly for redirecting RNA splicing particularly in therapeutic applications such as Duchenne muscular dystrophy (DMD). Covalent attachment of a cell-penetrating peptide helps to improve cell delivery of PNA. We have used a HeLa pLuc705 cell splicing redirection assay to develop a series of PNA internalization peptides (Pip) conjugated to an 18-mer PNA705 model oligonucleotide with higher activity compared to a PNA705 conjugate with a leading cell-penetrating peptide being developed for therapeutic use, (R-Ahx-R)4. We show that Pip–PNA705 conjugates are internalized in HeLa cells by an energy-dependent mechanism and that the predominant pathway of cell uptake of biologically active conjugate seems to be via clathrin-dependent endocytosis. In a mouse model of DMD, serum-stabilized Pip2a or Pip2b peptides conjugated to a 20-mer PNA (PNADMD) targeting the exon 23 mutation in the dystrophin gene showed strong exon-skipping activity in differentiated mdx mouse myotubes in culture in the absence of an added transfection agent at concentrations where naked PNADMD was inactive. Injection of Pip2a-PNADMD or Pip2b-PNADMD into the tibealis anterior muscles of mdx mice resulted in  3-fold higher numbers of dystrophin-positive fibres compared to naked PNADMD or (R-Ahx-R)4-PNADMD.
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