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Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein

A major rate-limiting step in nonviral gene delivery is the entry of nucleic acids across various membrane barriers and eventually into the nucleus where it must be transcribed. Cell-penetrating peptides and proteins are employed to generate formulations that overcome these challenges to facilitate...

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Autores principales: Jeyarajan, Sivakumar, Xavier, Jennifer, Rao, N Madhusudhana, Gopal, Vijaya
Formato: Texto
Lenguaje:English
Publicado: Dove Medical Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2962268/
https://www.ncbi.nlm.nih.gov/pubmed/21042418
http://dx.doi.org/10.2147/IJN.S13040
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author Jeyarajan, Sivakumar
Xavier, Jennifer
Rao, N Madhusudhana
Gopal, Vijaya
author_facet Jeyarajan, Sivakumar
Xavier, Jennifer
Rao, N Madhusudhana
Gopal, Vijaya
author_sort Jeyarajan, Sivakumar
collection PubMed
description A major rate-limiting step in nonviral gene delivery is the entry of nucleic acids across various membrane barriers and eventually into the nucleus where it must be transcribed. Cell-penetrating peptides and proteins are employed to generate formulations that overcome these challenges to facilitate DNA delivery into cells efficiently. However, these are limited by their inability to deliver nucleic acids selectively due to lack of specificity because they deliver to both cancer and normal cells. In this study, through modular design, we generated a recombinant fusion protein designated as Her-nuclear localization sequence (Her-NLS), where heregulin-α (Her), a targeting moiety, was cloned in frame with cationic NLS peptide to obtain a cell-specific targeting biomolecule for nucleic acid delivery. The heregulin-α(1) isoform possesses the epidermal growth factor-like domain and binds to HER2/3 heterodimers which are overexpressed in certain breast cancers. Purified recombinant Her-NLS fusion protein binds plasmid DNA and specifically transfects MDA-MB-453 cells overexpressing the epidermal growth factor receptors HER2/3 in vitro. The approach described would also permit replacement of heregulin ligand with other targeting moieties that would be suited to cell-specific nucleic acid delivery mediated via receptor-ligand interactions.
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spelling pubmed-29622682010-11-01 Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein Jeyarajan, Sivakumar Xavier, Jennifer Rao, N Madhusudhana Gopal, Vijaya Int J Nanomedicine Rapid Communication A major rate-limiting step in nonviral gene delivery is the entry of nucleic acids across various membrane barriers and eventually into the nucleus where it must be transcribed. Cell-penetrating peptides and proteins are employed to generate formulations that overcome these challenges to facilitate DNA delivery into cells efficiently. However, these are limited by their inability to deliver nucleic acids selectively due to lack of specificity because they deliver to both cancer and normal cells. In this study, through modular design, we generated a recombinant fusion protein designated as Her-nuclear localization sequence (Her-NLS), where heregulin-α (Her), a targeting moiety, was cloned in frame with cationic NLS peptide to obtain a cell-specific targeting biomolecule for nucleic acid delivery. The heregulin-α(1) isoform possesses the epidermal growth factor-like domain and binds to HER2/3 heterodimers which are overexpressed in certain breast cancers. Purified recombinant Her-NLS fusion protein binds plasmid DNA and specifically transfects MDA-MB-453 cells overexpressing the epidermal growth factor receptors HER2/3 in vitro. The approach described would also permit replacement of heregulin ligand with other targeting moieties that would be suited to cell-specific nucleic acid delivery mediated via receptor-ligand interactions. Dove Medical Press 2010-10-05 2010 /pmc/articles/PMC2962268/ /pubmed/21042418 http://dx.doi.org/10.2147/IJN.S13040 Text en © 2010 Jeyarajan et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article which permits unrestricted noncommercial use, provided the original work is properly cited.
spellingShingle Rapid Communication
Jeyarajan, Sivakumar
Xavier, Jennifer
Rao, N Madhusudhana
Gopal, Vijaya
Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title_full Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title_fullStr Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title_full_unstemmed Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title_short Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein
title_sort plasmid dna delivery into mda-mb-453 cells mediated by recombinant her-nls fusion protein
topic Rapid Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2962268/
https://www.ncbi.nlm.nih.gov/pubmed/21042418
http://dx.doi.org/10.2147/IJN.S13040
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