Highly efficient delivery of siRNA to a heart transplant model by a novel cell penetrating peptide-dsRNA binding domain
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Highly efficient delivery of siRNA to a heart transplant model by a novel cell penetrating peptide-dsRNA binding domain. / Li, Hua; Zheng, Xiangtao; Koren, Viktoria; Vashist, Yogesh Kumar; Tsui, Tung Yu.
In: INT J PHARMACEUT, Vol. 469, No. 1, 20.07.2014, p. 206-13.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Highly efficient delivery of siRNA to a heart transplant model by a novel cell penetrating peptide-dsRNA binding domain
AU - Li, Hua
AU - Zheng, Xiangtao
AU - Koren, Viktoria
AU - Vashist, Yogesh Kumar
AU - Tsui, Tung Yu
N1 - Copyright © 2014 Elsevier B.V. All rights reserved.
PY - 2014/7/20
Y1 - 2014/7/20
N2 - Small interfering RNAs (siRNAs) delivery remains a bottleneck for RNA interference (RNAi) - based therapies in the clinic. In the present study, a fusion protein with two cell-penetrating peptides (CPP), Hph1-Hph1, and a double-stranded RNA binding domain (dsRBD), was constructed for the siRNA delivery: dsRBD was designed to bind siRNA, and CPP would subsequently transport the dsRBD/siRNA complex into cells. We assessed the efficiency of the fusion protein, Hph1-Hph1-dsRBD, as a siRNA carrier. Calcium-condensed effects were assessed on GAPDH and green fluorescent protein (GFP) genes by western blot, real time polymerase chain reaction (RT-PCR), and flow cytometry analysis in vitro. Evaluations were also made in an in vivo heart transplantation model. The results demonstrated that the fusion protein, Hph1-Hph1-dsRBD, is highly efficient at delivering siRNA in vitro, and exhibits efficiency on GAPDH and GFP genes similar to or greater than lipofectamine. Interestingly, the calcium-condensed effects dramatically enhanced cellular uptake of the protein-siRNA complex. In vivo, Hph1-Hph1-dsRBD transferred and distributed ^ targeted siRNA throughout the whole mouse heart graft. Together, these results indicate that Hph1-Hph1-dsRBD has potential as an siRNA carrier for applications in the clinic or in biomedical research.
AB - Small interfering RNAs (siRNAs) delivery remains a bottleneck for RNA interference (RNAi) - based therapies in the clinic. In the present study, a fusion protein with two cell-penetrating peptides (CPP), Hph1-Hph1, and a double-stranded RNA binding domain (dsRBD), was constructed for the siRNA delivery: dsRBD was designed to bind siRNA, and CPP would subsequently transport the dsRBD/siRNA complex into cells. We assessed the efficiency of the fusion protein, Hph1-Hph1-dsRBD, as a siRNA carrier. Calcium-condensed effects were assessed on GAPDH and green fluorescent protein (GFP) genes by western blot, real time polymerase chain reaction (RT-PCR), and flow cytometry analysis in vitro. Evaluations were also made in an in vivo heart transplantation model. The results demonstrated that the fusion protein, Hph1-Hph1-dsRBD, is highly efficient at delivering siRNA in vitro, and exhibits efficiency on GAPDH and GFP genes similar to or greater than lipofectamine. Interestingly, the calcium-condensed effects dramatically enhanced cellular uptake of the protein-siRNA complex. In vivo, Hph1-Hph1-dsRBD transferred and distributed ^ targeted siRNA throughout the whole mouse heart graft. Together, these results indicate that Hph1-Hph1-dsRBD has potential as an siRNA carrier for applications in the clinic or in biomedical research.
KW - Animals
KW - Biological Transport
KW - Calcium
KW - Cell-Penetrating Peptides
KW - Gene Expression Regulation
KW - Gene Transfer Techniques
KW - Genes, Reporter
KW - Genetic Therapy
KW - Glyceraldehyde-3-Phosphate Dehydrogenases
KW - Green Fluorescent Proteins
KW - HeLa Cells
KW - Heart Transplantation
KW - Humans
KW - Male
KW - Mice, Inbred C57BL
KW - Peptide Fragments
KW - Protein Interaction Domains and Motifs
KW - RNA Interference
KW - RNA, Small Interfering
KW - RNA-Binding Proteins
KW - Recombinant Fusion Proteins
U2 - 10.1016/j.ijpharm.2014.04.050
DO - 10.1016/j.ijpharm.2014.04.050
M3 - SCORING: Journal article
C2 - 24768403
VL - 469
SP - 206
EP - 213
JO - INT J PHARMACEUT
JF - INT J PHARMACEUT
SN - 0378-5173
IS - 1
ER -