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A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin
Nonregenerative anemia due to chronic renal failure is a common problem in domestic cats. Unfortunately, administration of recombinant human erythropoietin often only improves anemia temporarily due to antibody development. In this in vitro study, feline erythropoietin cDNA was cloned from feline re...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3445592/ https://www.ncbi.nlm.nih.gov/pubmed/23028782 http://dx.doi.org/10.1371/journal.pone.0045099 |
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author | Vapniarsky, Natalia Lame, Michael McDonnel, Samantha Murphy, Brian |
author_facet | Vapniarsky, Natalia Lame, Michael McDonnel, Samantha Murphy, Brian |
author_sort | Vapniarsky, Natalia |
collection | PubMed |
description | Nonregenerative anemia due to chronic renal failure is a common problem in domestic cats. Unfortunately, administration of recombinant human erythropoietin often only improves anemia temporarily due to antibody development. In this in vitro study, feline erythropoietin cDNA was cloned from feline renal tissue and utilized in the construction of a replication-defective lentiviral vector. The native recombinant feline erythropoietin (rfEPO) sequence was confirmed by sequencing. Upon viral vector infection of human 293H cells, Crandall Renal Feline Kidney cell line and primary feline peripheral blood mononuclear cells, bioactive rfEPO protein was produced. The presence of cellular rfEPO cDNA was confirmed by standard PCR, production of abundant rfEPO mRNA was confirmed by real-time PCR, and secretion of rfEPO protein was demonstrated by Western blot analyses, while rfEPO protein bioactivity was confirmed via an MTT proliferation bioassay. This in vitro study demonstrates the feasibility of a replication-defective lentiviral vector delivery system for the in vitro production of biologically active feline erythropoietin. Anemic cats with chronic renal failure represent a potential in vivo application of a lentiviral gene therapy system. |
format | Online Article Text |
id | pubmed-3445592 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-34455922012-10-01 A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin Vapniarsky, Natalia Lame, Michael McDonnel, Samantha Murphy, Brian PLoS One Research Article Nonregenerative anemia due to chronic renal failure is a common problem in domestic cats. Unfortunately, administration of recombinant human erythropoietin often only improves anemia temporarily due to antibody development. In this in vitro study, feline erythropoietin cDNA was cloned from feline renal tissue and utilized in the construction of a replication-defective lentiviral vector. The native recombinant feline erythropoietin (rfEPO) sequence was confirmed by sequencing. Upon viral vector infection of human 293H cells, Crandall Renal Feline Kidney cell line and primary feline peripheral blood mononuclear cells, bioactive rfEPO protein was produced. The presence of cellular rfEPO cDNA was confirmed by standard PCR, production of abundant rfEPO mRNA was confirmed by real-time PCR, and secretion of rfEPO protein was demonstrated by Western blot analyses, while rfEPO protein bioactivity was confirmed via an MTT proliferation bioassay. This in vitro study demonstrates the feasibility of a replication-defective lentiviral vector delivery system for the in vitro production of biologically active feline erythropoietin. Anemic cats with chronic renal failure represent a potential in vivo application of a lentiviral gene therapy system. Public Library of Science 2012-09-18 /pmc/articles/PMC3445592/ /pubmed/23028782 http://dx.doi.org/10.1371/journal.pone.0045099 Text en © 2012 Vapniarsky et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Vapniarsky, Natalia Lame, Michael McDonnel, Samantha Murphy, Brian A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title | A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title_full | A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title_fullStr | A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title_full_unstemmed | A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title_short | A Lentiviral Gene Therapy Strategy for the In Vitro Production of Feline Erythropoietin |
title_sort | lentiviral gene therapy strategy for the in vitro production of feline erythropoietin |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3445592/ https://www.ncbi.nlm.nih.gov/pubmed/23028782 http://dx.doi.org/10.1371/journal.pone.0045099 |
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