Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides

Bo Lou; Chun Yin Jerry Lau; Wim E Hennink; Enrico Mastrobattista

doi:10.1007/978-1-0716-1617-8_21

Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides

Bo Lou, Chun Yin Jerry Lau, Wim E Hennink, Enrico Mastrobattista^*

^*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

Abstract

Successful delivery of mRNA into the cytosol of professional antigen-presenting cells (APCs) poses one of the biggest challenges in developing effective mRNA vaccines to treat various cancers and viral infectious diseases. However, most polymeric mRNA delivery systems fail to transfect APCs. We have discovered that decoration of pH-sensitive endosome-disruptive GALA peptides on the surface of mRNA polyplexes leads to efficient targeting and transfection of APCs. GALA peptides not only enhance specific uptake in APCs through binding to sialic acid moieties, they also facilitate the endosomal escape of mRNA especially in dendritic cells (DCs). Here, we describe in detail the production of stabilized mRNA polyplexes post-conjugated with GALA peptides via copper-free click chemistry. Methods described here include the synthesis and purification of GALA peptides and its conjugation to mRNA polyplexes.

Original language	English
Title of host publication	Peptide Conjugation
Subtitle of host publication	Methods and Protocols
Editors	Waleed M. Hussein, Rachel J. Stephenson, Istvan Toth
Place of Publication	New York, NY
Publisher	Humana Press
Pages	275-286
Number of pages	12
Edition	1
ISBN (Electronic)	978-1-0716-1617-8
ISBN (Print)	978-1-0716-1616-1, 978-1-0716-1619-2
DOIs	https://doi.org/10.1007/978-1-0716-1617-8_21
Publication status	Published - 14 Aug 2021

Publication series

Name	Methods in Molecular Biology
Publisher	Humana
Volume	2355
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

Keywords

Endosomes
Peptides
RNA, Messenger/genetics
Transfection
mRNA Vaccines

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/978-1-0716-1617-8_21

Lou2021_Protocol_PreparationOfMRNAPolyplexesWitFinal published version, 313 KBLicence: Taverne

Cite this

Lou, B., Lau, C. Y. J., Hennink, W. E., & Mastrobattista, E. (2021). Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides. In W. M. Hussein, R. J. Stephenson, & I. Toth (Eds.), Peptide Conjugation: Methods and Protocols (1 ed., pp. 275-286). (Methods in Molecular Biology; Vol. 2355). Humana Press. https://doi.org/10.1007/978-1-0716-1617-8_21

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abstract = "Successful delivery of mRNA into the cytosol of professional antigen-presenting cells (APCs) poses one of the biggest challenges in developing effective mRNA vaccines to treat various cancers and viral infectious diseases. However, most polymeric mRNA delivery systems fail to transfect APCs. We have discovered that decoration of pH-sensitive endosome-disruptive GALA peptides on the surface of mRNA polyplexes leads to efficient targeting and transfection of APCs. GALA peptides not only enhance specific uptake in APCs through binding to sialic acid moieties, they also facilitate the endosomal escape of mRNA especially in dendritic cells (DCs). Here, we describe in detail the production of stabilized mRNA polyplexes post-conjugated with GALA peptides via copper-free click chemistry. Methods described here include the synthesis and purification of GALA peptides and its conjugation to mRNA polyplexes.",

keywords = "Endosomes, Peptides, RNA, Messenger/genetics, Transfection, mRNA Vaccines",

author = "Bo Lou and Lau, {Chun Yin Jerry} and Hennink, {Wim E} and Enrico Mastrobattista",

note = "{\textcopyright} 2021. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",

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Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides. / Lou, Bo; Lau, Chun Yin Jerry; Hennink, Wim E et al.
Peptide Conjugation: Methods and Protocols. ed. / Waleed M. Hussein; Rachel J. Stephenson; Istvan Toth. 1. ed. New York, NY: Humana Press, 2021. p. 275-286 (Methods in Molecular Biology; Vol. 2355).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

TY - CHAP

T1 - Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides

AU - Lou, Bo

AU - Lau, Chun Yin Jerry

AU - Hennink, Wim E

AU - Mastrobattista, Enrico

PY - 2021/8/14

Y1 - 2021/8/14

N2 - Successful delivery of mRNA into the cytosol of professional antigen-presenting cells (APCs) poses one of the biggest challenges in developing effective mRNA vaccines to treat various cancers and viral infectious diseases. However, most polymeric mRNA delivery systems fail to transfect APCs. We have discovered that decoration of pH-sensitive endosome-disruptive GALA peptides on the surface of mRNA polyplexes leads to efficient targeting and transfection of APCs. GALA peptides not only enhance specific uptake in APCs through binding to sialic acid moieties, they also facilitate the endosomal escape of mRNA especially in dendritic cells (DCs). Here, we describe in detail the production of stabilized mRNA polyplexes post-conjugated with GALA peptides via copper-free click chemistry. Methods described here include the synthesis and purification of GALA peptides and its conjugation to mRNA polyplexes.

AB - Successful delivery of mRNA into the cytosol of professional antigen-presenting cells (APCs) poses one of the biggest challenges in developing effective mRNA vaccines to treat various cancers and viral infectious diseases. However, most polymeric mRNA delivery systems fail to transfect APCs. We have discovered that decoration of pH-sensitive endosome-disruptive GALA peptides on the surface of mRNA polyplexes leads to efficient targeting and transfection of APCs. GALA peptides not only enhance specific uptake in APCs through binding to sialic acid moieties, they also facilitate the endosomal escape of mRNA especially in dendritic cells (DCs). Here, we describe in detail the production of stabilized mRNA polyplexes post-conjugated with GALA peptides via copper-free click chemistry. Methods described here include the synthesis and purification of GALA peptides and its conjugation to mRNA polyplexes.

KW - Endosomes

KW - Peptides

KW - RNA, Messenger/genetics

KW - Transfection

KW - mRNA Vaccines

U2 - 10.1007/978-1-0716-1617-8_21

DO - 10.1007/978-1-0716-1617-8_21

M3 - Chapter

C2 - 34386965

SN - 978-1-0716-1616-1

SN - 978-1-0716-1619-2

T3 - Methods in Molecular Biology

SP - 275

EP - 286

BT - Peptide Conjugation

A2 - Hussein, Waleed M.

A2 - Stephenson, Rachel J.

A2 - Toth, Istvan

PB - Humana Press

CY - New York, NY

ER -

Preparation of mRNA Polyplexes with Post-conjugated Endosome-Disruptive Peptides

Abstract

Publication series

Bibliographical note

Keywords

UN SDGs

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Cite this