Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method

Martina Tuttolomondo; Henrik J. Ditzel

doi:10.1007/978-1-0716-1298-9_4

Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method

Martina Tuttolomondo^*
, Henrik J. Ditzel

^*Kontaktforfatter

KI, OUH, Forskningsenhed for Onkologi (Odense)

Publikation: Kapitel i bog/rapport/konference-proceeding › Kapitel i bog › Forskning › peer review

Abstract

Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

Originalsprog	Engelsk
Titel	Design and Delivery of SiRNA Therapeutics
Redaktører	Henrik J. Ditzel, Martina Tuttolomondo, Sakari Kauppinen
Vol/bind	2282
Forlag	Humana Press
Publikationsdato	2021
Sider	43-56
ISBN (Trykt)	978-1-0716-1297-2
ISBN (Elektronisk)	978-1-0716-1298-9
DOI	https://doi.org/10.1007/978-1-0716-1298-9_4
Status	Udgivet - 2021

Navn	Methods in Molecular Biology
Vol/bind	2282
ISSN	1064-3745

Bibliografisk note

Funding Information:
We gratefully acknowledge funding support from A Race Agaisnt Breast Cancer and the A.P. Moeller Foundation. Moreover, we acknowledge Dr. Bartosz Pilecki from the Department of Cancer and Inflammation Research, University of Southern Denmark, for providing the mouse serum.

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2021.

Finansiering

We gratefully acknowledge funding support from A Race Agaisnt Breast Cancer and the A.P. Moeller Foundation. Moreover, we acknowledge Dr. Bartosz Pilecki from the Department of Cancer and Inflammation Research, University of Southern Denmark, for providing the mouse serum.

Dokumenter og links

10.1007/978-1-0716-1298-9_4

SDU link

Tjek adgangen til materialet i Syddansk Universitetsbiblioteks søgemaskine

Citationsformater

Tuttolomondo, Martina ; Ditzel, Henrik J. / Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method. Design and Delivery of SiRNA Therapeutics. red. / Henrik J. Ditzel ; Martina Tuttolomondo ; Sakari Kauppinen. Bind 2282 Humana Press, 2021. s. 43-56 (Methods in Molecular Biology, Bind 2282).

@inbook{f66735eb7f82404d91bd073c31f6ed7f,

title = "Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method",

abstract = "Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast F{\"o}rster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.",

keywords = "Agarose gel electrophoresis, F{\"o}rster resonance energy transfer, RNA interference, Serum protein corona, siRNA stability, Small interfering RNA",

author = "Martina Tuttolomondo and Ditzel, \{Henrik J.\}",

note = "Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2021.",

year = "2021",

doi = "10.1007/978-1-0716-1298-9\_4",

language = "English",

isbn = " 978-1-0716-1297-2",

volume = "2282",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "43--56",

editor = "Ditzel, \{Henrik J.\} and \{ Tuttolomondo\}, Martina and Sakari Kauppinen",

booktitle = "Design and Delivery of SiRNA Therapeutics",

address = "United States",

}

Tuttolomondo, M & Ditzel, HJ 2021, Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method. i HJ Ditzel, M Tuttolomondo & S Kauppinen (red), Design and Delivery of SiRNA Therapeutics. bind 2282, Humana Press, Methods in Molecular Biology, bind 2282, s. 43-56. https://doi.org/10.1007/978-1-0716-1298-9_4

Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method. / Tuttolomondo, Martina ; Ditzel, Henrik J.
Design and Delivery of SiRNA Therapeutics. red. / Henrik J. Ditzel; Martina Tuttolomondo; Sakari Kauppinen. Bind 2282 Humana Press, 2021. s. 43-56 (Methods in Molecular Biology, Bind 2282).

Publikation: Kapitel i bog/rapport/konference-proceeding › Kapitel i bog › Forskning › peer review

TY - CHAP

T1 - Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method

AU - Tuttolomondo, Martina

AU - Ditzel, Henrik J.

N1 - Publisher Copyright: © Springer Science+Business Media, LLC, part of Springer Nature 2021.

PY - 2021

Y1 - 2021

N2 - Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

AB - Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

KW - Agarose gel electrophoresis

KW - Förster resonance energy transfer

KW - RNA interference

KW - Serum protein corona

KW - siRNA stability

KW - Small interfering RNA

U2 - 10.1007/978-1-0716-1298-9_4

DO - 10.1007/978-1-0716-1298-9_4

M3 - Book chapter

C2 - 33928569

AN - SCOPUS:85105159646

SN - 978-1-0716-1297-2

VL - 2282

T3 - Methods in Molecular Biology

SP - 43

EP - 56

BT - Design and Delivery of SiRNA Therapeutics

A2 - Ditzel, Henrik J.

A2 - Tuttolomondo, Martina

A2 - Kauppinen, Sakari

PB - Humana Press

ER -

Evaluation of siRNA stability and interaction with serum components using an agarose gel-based single-molecule FRET labeling method

Abstract

Bibliografisk note

Finansiering

Dokumenter og links

SDU link

Fingeraftryk

Citationsformater