Open AccessInternalization of Locked Nucleic Acids/DNA Hybrid Oligomers into Escherichia coli
Author(s) -
Germán M. Traglia,
Cornelia Sala,
Juan I. Fuxman Bass,
Alfonso SolerBistué,
Ãngeles Zorreguieta,
María Soledad Ramírez,
Marcelo E. Tolmasky
Publication year - 2012
Publication title -
bioresearch open access
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.457
H-Index - 26
eISSN - 2164-7860
pISSN - 2164-7844
DOI - 10.1089/biores.2012.0257
Subject(s) - nucleic acid , escherichia coli , dna , cytoplasm , alexa fluor , internalization , microbiology and biotechnology , flow cytometry , locked nucleic acid , confocal microscopy , biology , chemistry , oligonucleotide , biochemistry , cell , fluorescence , gene , quantum mechanics , physics
Delivery inside the cells is essential for practical application of antisense technologies. The hybrid locked nucleic acid (LNA)/DNA CAAGTACTGTTCCACCA (LNA residues are underlined) was labeled by conjugation to Alexa Fluor 488 (fLNA/DNA) and tested to determine its ability to penetrate Escherichia coli cells and reach the cytoplasm. Flow cytometry analysis showed that the fLNA/DNA was associated with 14% of cells from a stationary phase culture, while association with a labeled isosequential oligodeoxynucleotide was negligible. Laser scanning confocal microscopy confirmed that the fLNA/DNA was located inside the cytoplasm.
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