Rapid Delivery of Nanobodies/VH Hs into Living Cells via Expressing In Vitro -Transcribed mRNA

Molecular Therapy. Methods & Clinical Development
Xuechen ZhouWei Xie

Abstract

Intracellular antigen labeling and manipulation by antibodies have been long-thought goals in the field of cell research and therapy. However, a central limitation for this application is that antibodies are not able to penetrate into the cytosol of living cells. Taking advantages of small sizes and unique structures of the single-domain antibodies, here, we presented a novel approach to rapidly deliver the nanobody/variable domain of heavy chain of heavy-chain antibody (VHH) into living cells via introducing its coding mRNA, which was generated by in vitro transcription. We demonstrated that actin-green fluorescent proteins (GFP) and Golgi-GFP can be recognized by the anti-GFP nanobody/VHH, vimentin can be recognized by the anti-vimentin nanobody/VHH, and histone deacetylase 6 (HDAC6) can be recognized by the anti-HDAC6 nanobody/VHH, respectively. We found that the anti-GFP nanobody expressed via in vitro-transcribed (IVT) mRNA can be detected in 3 h and degraded in 48 h after transfection, whereas the nanobody expressed via plasmid DNA, was not detected until 24 h after transfection. As a result, it is effective in delivering the nanobody through expressing the nanobody/VHH in living cells from its coding mRNA.

References

Jun 3, 1993·Nature·C Hamers-CastermanR Hamers
Dec 29, 1998·The Journal of Cell Biology·J A JohnstonR R Kopito
Jun 30, 2000·Current Opinion in Cell Biology·A Prochiantz
Jun 28, 2005·Current Opinion in Chemical Biology·Martin Stocks
Oct 25, 2006·Nature Methods·Ulrich RothbauerHeinrich Leonhardt
Jun 7, 2007·Nature Reviews. Molecular Cell Biology·Harald HerrmannUeli Aebi
Jun 10, 2008·Nature Methods·Julia RiedlRoland Wedlich-Soldner
Nov 13, 2008·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Lioudmila V SharovaMinoru S H Ko
Sep 3, 2009·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Martin A CheeverLynn M Matrisian
Apr 24, 2010·Nature Reviews. Immunology·Andrew C Chan, Paul J Carter
Apr 29, 2010·Nature Reviews. Immunology·Olive Leavy
Oct 15, 2010·Protein Science : a Publication of the Protein Society·Marta H KubalaBrett M Collins
Nov 26, 2010·MAbs·Andrea L J MarschallStefan Dübel
Jan 11, 2011·Nature Biotechnology·Michael S D KormannCarsten Rudolph
Apr 28, 2012·Cellular and Molecular Life Sciences : CMLS·Lucy W BarrettSteve D Wilton
Jun 20, 2012·Current Protein & Peptide Science·Xavier PichonAnne E E Willis
Sep 23, 2014·Nature Reviews. Drug Discovery·Ugur SahinÖzlem Türeci
Jul 4, 2015·Histochemistry and Cell Biology·Hiroshi KimuraYuko Sato
Feb 4, 2016·International Journal of Molecular Sciences·Mie KristensenHanne Mørck Nielsen
Feb 11, 2016·The New England Journal of Medicine·Flora PeyvandiUNKNOWN TITAN Investigators
Feb 26, 2016·International Journal of Molecular Sciences·Ana DincaMichael T Chin
Jul 30, 2017·Nature Chemistry·Macarena Sánchez-NavarroErnest Giralt
Aug 20, 2017·Protein & Cell·William R Strohl
Sep 5, 2017·Frontiers in Immunology·Iva TrenevskaAlison H Banham
Dec 2, 2017·Nature Reviews. Drug Discovery·Paul J Carter, Greg A Lazar
Jan 13, 2018·Nature Reviews. Drug Discovery·Norbert PardiDrew Weissman
May 12, 2018·Science·Mitch Leslie
May 25, 2018·Journal of the American Chemical Society·Matthew G CostalesMatthew D Disney
Sep 8, 2018·Nature Reviews. Drug Discovery·Fernando PastorIgnacio Melero
Oct 31, 2018·Nature Communications·Nuphar VeigaDan Peer

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Citations

Dec 3, 2020·American Journal of Physiology. Cell Physiology·Jan Gettemans, Brian De Dobbelaer
Dec 17, 2020·Biomolecules·Eline SoetensXavier Saelens
Dec 24, 2020·Biomolecules·Marco ErreniAndrea Doni

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Methods Mentioned

BETA
transfection
PCR
in vitro transcription
transfections

Software Mentioned

ImageJ
GraphPad Prism
GENEWIZ

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