Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors

Oliver Bogen; Dionéia Araldi; Anatol Sucher; Kord Kober; Peter T Ohara; Jon D Levine

doi:10.1177/17448069241230419

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Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors

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Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors

Oliver Bogen, Dionéia Araldi, Anatol Sucher, Kord Kober, Peter T Ohara and Jon D Levine

Molecular pain, Vol.20, p.17448069241230419

01/01/2024

DOI: 10.1177/17448069241230419

PMID: 38246917

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Abstract

In vivo analysis of protein function in nociceptor subpopulations using antisense oligonucleotides and short interfering RNAs is limited by their non-selective cellular uptake. To address the need for selective transfection methods, we covalently linked isolectin B4 (IB4) to streptavidin and analyzed whether it could be used to study protein function in IB4(+)-nociceptors. Rats treated intrathecally with IB4-conjugated streptavidin complexed with biotinylated antisense oligonucleotides for protein kinase C epsilon (PKCε) mRNA were found to have: (a) less PKCε in dorsal root ganglia (DRG), (b) reduced PKCε expression in IB4(+) but not IB4(–) DRG neurons, and (c) fewer transcripts of the PKCε gene in the DRG. This knockdown in PKCε expression in IB4(+) DRG neurons is sufficient to reverse hyperalgesic priming, a rodent model of chronic pain that is dependent on PKCε in IB4(+)-nociceptors. These results establish that IB4-streptavidin can be used to study protein function in a defined subpopulation of nociceptive C-fiber afferents.

Details

Title: Subtitle: Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors
Creators: Oliver Bogen - University of California, San Francisco
Dionéia Araldi - University of California, San Francisco
Anatol Sucher - University of California, San Francisco
Kord Kober - University of California, San Francisco
Peter T Ohara - University of California, San Francisco
Jon D Levine - University of California, San Francisco
Resource Type: Journal article
Publication Details: Molecular pain, Vol.20, p.17448069241230419
DOI: 10.1177/17448069241230419
PMID: 38246917
NLM abbreviation: Mol Pain
ISSN: 1744-8069
eISSN: 1744-8069
Publisher: SAGE Publications
Grant note: R01CA250017 / ; R01AR075334 / ;
Language: English
Date published: 01/01/2024
Academic Unit: Neuroscience and Pharmacology
Record Identifier: 9985177944802771

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