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2017 ; 1521
(ä): 139-152
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Exosomes-Based Gene Therapy for MicroRNA Delivery
#MMPMID27910046
Mathiyalagan P
; Sahoo S
Methods Mol Biol
2017[]; 1521
(ä): 139-152
PMID27910046
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Despite recent advances in scientific knowledge and clinical practice,
cardiovascular disease management and treatment remain a major burden. While
several treatment strategies using drugs and surgeries are being developed for
cardiovascular manifestations, gene-based therapies hold significant promise.
Recent findings from our laboratory unveiled a novel mechanism that exosomes,
secreted nanovesicles from stem cells, mediate cardiac repair via transferring
their unique repertoire of microRNAs (miRNA) to recipient cells in the heart.
Exosomes, unlike other vectors for gene delivery, present unique advantages such
that exosomes are a cell-free natural system for ferrying RNA between cells,
robust exosomal membrane can protect the RNA/gene of interest from digestion, and
exosomes are rapidly taken up by target cells making them a more efficient
vehicle for gene delivery. Here, we describe a stepwise protocol developed in our
laboratory for generating exosomes from human CD34(+) stem cells that carry
exogenously applied Cy3 dye-labeled pre-miR miRNA precursors. We demonstrate that
human CD34(+) stem cell exosomes can rigorously enter into recipient cells and
deliver Cy3 dye-labeled pre-miR miRNA precursors to regulate gene expression.
Identification of key molecular targets to treat disease conditions is the
foremost critical step and the novel approach presented here to generate exosomes
carrying exogenous genetic information offers a valuable clinical tool for more
effective treatment strategies.
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