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Identification of a cardiac specific protein transduction domain by in Vivo biopanning using a M13 phage peptide display library in mice

Zahid, M and Phillips, BE and Albers, SM and Giannoukakis, N and Watkins, SC and Robbins, PD (2010) Identification of a cardiac specific protein transduction domain by in Vivo biopanning using a M13 phage peptide display library in mice. PLoS ONE, 5 (8).

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Background: A peptide able to transduce cardiac tissue specifically, delivering cargoes to the heart, would be of significant therapeutic potential for delivery of small molecules, proteins and nucleic acids. In order to identify peptide(s) able to transduce heart tissue, biopanning was performed in cell culture and in vivo with a M13 phage peptide display library. Methods and Results: A cardiomyoblast cell line, H9C2, was incubated with a M13 phage 12 amino acid peptide display library. Internalized phage was recovered, amplified and then subjected to a total of three rounds of in vivo biopanning where infectious phage was isolated from cardiac tissue following intravenous injection. After the third round, 60% of sequenced plaques carried the peptide sequence APWHLSSQYSRT, termed cardiac targeting peptide (CTP). We demonstrate that CTP was able to transduce cardiomyocytes functionally in culture in a concentration and cell-type dependent manner. Mice injected with CTP showed significant transduction of heart tissue with minimal uptake by lung and kidney capillaries, and no uptake in liver, skeletal muscle, spleen or brain. The level of heart transduction by CTP also was greater than with a cationic transduction domain. Conclusions: Biopanning using a peptide phage display library identified a peptide able to transduce heart tissue in vivo efficiently and specifically. CTP could be used to deliver therapeutic peptides, proteins and nucleic acid specifically to the heart. © 2010 Zahid et al.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Zahid, Mmaz7@pitt.eduMAZ7
Phillips, BE
Albers, SM
Giannoukakis, Nngiann1@pitt.eduNGIANN1
Watkins, SCsimon.watkins@pitt.eduSWATKINS
Robbins, PDprobb@pitt.eduPROBB
ContributionContributors NameEmailPitt UsernameORCID
Centers: Other Centers, Institutes, Offices, or Units > Center for Biologic Imaging
Date: 19 October 2010
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 5
Number: 8
DOI or Unique Handle: 10.1371/journal.pone.0012252
Refereed: Yes
MeSH Headings: Amino Acid Sequence; Animals; Bacteriophage M13; Biological Transport; Cell Culture Techniques; Cell Line; Female; Humans; Membrane Transport Proteins--chemistry; Membrane Transport Proteins--metabolism; Mice; Microscopy, Confocal; Myoblasts, Cardiac--cytology; Myoblasts, Cardiac--metabolism; Myocardium--cytology; Myocardium--metabolism; Myocytes, Cardiac--cytology; Myocytes, Cardiac--metabolism; NF-kappa B--metabolism; Oligopeptides--chemistry; Oligopeptides--metabolism; Organ Specificity; Peptide Library; Protein Structure, Tertiary; Protein-Serine-Threonine Kinases--metabolism; Signal Transduction
Other ID: NLM PMC2923200
PubMed Central ID: PMC2923200
PubMed ID: 20808875
Date Deposited: 15 Aug 2012 17:56
Last Modified: 02 Feb 2019 16:56


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