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CLIP6-PNA-Peptide Conjugates: Non-Endosomal Delivery of Splice Switching Oligonucleotides
journal contribution
posted on 2017-12-06, 20:40 authored by Terese Soudah, Maxim Mogilevsky, Rotem Karni, Eylon YavinEfficient delivery of oligonucleotides
still remains a challenge
in the field of oligonucleotide based therapy. Peptide nucleic acid
(PNA), a DNA analogue that is typically synthesized by solid phase
peptide chemistry, has been conjugated to a variety of cell penetrating
peptides (CPP) as a means of improving its cellular uptake. These
CPPs typically deliver their cargoes into cells by an endosomal-dependent
mechanism resulting in lower bioavailability of the cargo. Herein,
we designed and synthesized PNA–peptide conjugates as splice
switching oligonucleotides (SSO) targeting the Mnk2 gene, a therapeutic
target in cancer. In humans, the MKNK2 gene, is alternatively spliced,
generating isoforms with opposite biological activities: Mnk2a and
Mnk2b. It was found that the Mnk2a isoform is down-regulated in breast,
lung, brain, and colon tumors and is a tumor suppressor, whereas MnK2b
is oncogenic. We have designed and synthesized PNAs that were conjugated
to either of the following peptides: a nuclear localization sequence
(NLS) or a cytosol localizing internalization peptide (CLIP6). CLIP6-PNA
demonstrates effective cellular uptake and exclusively employs a nonendosomal
mechanism to cross the cellular membranes of glioblastoma cells (U87).
Simple incubation of PNA–peptide conjugates in human glioblastoma
cells up-regulates the Mnk2a isoform leading to cancer cell death.
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localization sequencecancer cell deathCPPglioblastoma cells up-regulatesuptakeendosomal-dependent mechanismSplice Switching Oligonucleotides Efficient deliveryglioblastoma cellsMKNK 2 genephase peptide chemistrycargoMnK 2btumor suppressornonendosomal mechanismNon-Endosomal DeliveryoligonucleotideMnk 2a isoformMnk 2 geneSSOcytosol localizing internalization peptideDNA analogueMnk 2aMnk 2bcolon tumorsconjugateCLIP 6-PNA ConjugatesNLS
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