Efficient delivery of miR-122 to regulate cholesterol metabolism using a non-covalent peptide-based strategy.
MicroRNAs (miRNAs) are small non-coding RNAs that are important in the pathogenesis of multiple diseases and, therefore, may represent a novel class of targets for therapeutic intervention. However, like the majority of oligonucleotide-based strategies, there are obstacles to their clinical application, including poor cellular uptake due to the low permeability of the cell membrane to negatively charged molecules. MPG is a 27-residue peptide vector which contains a hydrophobic domain derived from the fusion sequence of HIV-1 gp41 and a hydrophilic domain derived from the nuclear localization sequence of SV40 T-antigen. MPG is one of the most promising tools for the non-invasive cellular import of oligonucleotides and analogs. In the present study, a non-covalent peptide-based strategy was used for the efficient delivery of the miRNA-122 (miR-122) mimic and inhibitor into mouse liver cell lines, mouse primary hepatocytes and C. elegans, without any associated cytotoxicity. Moreover, high-performance liquid chromatography analysis determined that MPG and MPGΔNLS delivered the miR-122 mimic and inhibitor into mouse liver cells and effectively regulated cholesterol levels. The results demonstrated that MPG family members may be used for the efficient delivery of miR-122 to regulate cholesterol metabolism, and that this cell-penetrating peptide-based technology may be beneficial for further biological applications of RNA therapeutics in vivo.
Center for Gene Diagnosis, Zhongnan Hospital, Wuhan University, Wuhan, Hubei 430071, P.R. China.
Lilin
L
Wang
Wei
W
Tang
Shirong
S
Yan
Long
L
Zhou
Tao
T
Shen
Xiuqing
X
Huang
Lin
L
Dou
Mo
M
Wang
Songlin
S
Yu
Jian
J
Li
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000166:cytology / Q000378:metabolism*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism*; D015700:HIV Envelope Protein gp41 / Q000235:genetics / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics*; D010446:Peptide Fragments / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000952:Antigens, Polyomavirus Transforming; D015700:HIV Envelope Protein gp41; D035683:MicroRNAs; C544354:Mirn122 microRNA, mouse; D010446:Peptide Fragments; D012333:RNA, Messenger; D002784:Cholesterol