title: A peptide nucleic acid (PNA)-mediated polymerase chain reaction clamping allows the selective inhibition of the ERVWE1 gene amplification.
abstract: A new peptide nucleic acid (PNA) mediated QPCR technique for the detection and quantification of the Multiple Sclerosis-Associated Retrovirus (MSRV) belonging to the human endogenous retrovirus-W (HERV-W) family has been developed. The assay utilizes a PNA probe which is fully complementary to the ERVWE1 sequence, another member of the HERV-W family which is closely related to MSRV. Due to its excellent affinity to a completely matched template, PNA probe selectively blocks the amplification of ERVWE1 thus allowing the specific and exclusive detection and quantification of the MSRV as PNA does not interfere with the amplification of MSRV. Using this newly developed method we found that MSRV is predominantly expressed over ERWVE1 in astrocyte-derived U-87 MG cell line but not in human umbilical vein endothelial cells or human placental cells.
authors.affiliation: Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland. Electronic address: gmachnik@sum.edu.pl.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Machnik
authors.affiliation: Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Łabuzek
authors.affiliation: Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland.
authors.forename: Estera
authors.initials: E
authors.lastname: Skudrzyk
authors.affiliation: Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-952 Gdańsk, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Rekowski
authors.affiliation: Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-952 Gdańsk, Poland.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Ruczyński
authors.affiliation: Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-952 Gdańsk, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Wojciechowska
authors.affiliation: Faculty of Chemistry, University of Gdańsk, Wita Stwosza 63, 80-952 Gdańsk, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Mucha
authors.affiliation: Department of Neurology, Henry Ford Hospital, 2799 West Grand Boulevard, Detroit, MI 48202, USA.
authors.forename: Shailendra
authors.initials: S
authors.lastname: Giri
authors.affiliation: Department of Pharmacology, Medical University of Silesia, Medyków 18, 40-752 Katowice, Poland.
authors.forename: Bogusław
authors.initials: B
authors.lastname: Okopień
mesh_terms: D001483:Base Sequence; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D020077:Endogenous Retroviruses / Q000235:genetics; D005260:Female; D005784:Gene Amplification; D015686:Gene Products, env / Q000235:genetics*; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000235:genetics*; D010920:Placenta / Q000166:cytology / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics*; D015203:Reproducibility of Results; D012689:Sequence Homology, Nucleic Acid
chemical_list: D015686:Gene Products, env; D020135:Peptide Nucleic Acids; D011257:Pregnancy Proteins; C405335:syncytin
keywords: ERVWE1; Endogenous retroviruses; Human; Mutation detection; Peptide nucleic acids (PNAs); Syncytin-1 glycoprotein