Transport of oligonucleotides across natural and model membranes

Vlassov, V. V.; Blakireva, Larissa A.; Yakubov, L. A.

doi:10.1016/0304-4157(94)90001-9

Cited by 89 publications

(34 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The other is internalization by receptor-mediated endocytosis. Somatic cells have been reported to express a variety of DNA receptor candidates, 25,32,33,[35][36][37][38] none of which showed to be uniquely essential for DNA uptake. Specifically, receptors mediating DNA internalization in human B lymphocytes are poorly understood.…”

Section: Discussionmentioning

confidence: 99%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

View full text Add to dashboard Cite

DNA can cross the cell membrane by natural means, but the functional relevance of this phenomenon has not been fully elucidated. Here, we analyzed spontaneous transgenesis of human B cells using plasmid DNA coding for a functional immunoglobulin (Ig) heavy chain gene under the control of a B-cell-specific promoter. Using polymerase chain reaction (PCR), reverse transcriptase-PCR, and flow cytometry in combination, spontaneous transgenesis was documented in Burkitt's lymphoma cell lines, Epstein-Barr virus-transformed cell lines, and peripheral blood B lymphocytes of the mature naïve phenotype (IgM þ /IgD þ /CD27 À ). By immunoelectron microscopy, the internalized DNA was seen in the lysosomes/late endosomes and in the cytosol proximal to the nucleus. Importantly, spontaneously transgenic B cells processed and presented to major histocompatibility complex (MHC)-restricted T lymphocytes a peptide expressed in the transgenic product. This is the first demonstration that primary B lymphocytes possess a program for the spontaneous internalization of DNA, which in turn imparts the cell with new immunological functions. As spontaneous transgenesis is obtained using a nonviral vector, does not require prior cell activation, and is not associated with chromosomal integration, the findings reported here open new possibilities for genetic manipulations of mature naïve B lymphocytes for therapy and vaccination.

show abstract

Section: Discussionmentioning

confidence: 99%

Spontaneous transgenesis of human B lymphocytes

et al. 2003

View full text Add to dashboard Cite

show abstract

“…Oligonucleotides are polyanionic species that are internalized in cells, probably by receptor-mediated endocytosis (37). They are likely to interact with many biomolecules within the cell and also in the extracellular membrane by virtue of both their charge and their shape, as well as sequence-specific interactions with nucleic acids.…”

Section: Discussionmentioning

confidence: 99%

Antiproliferative Activity of G-rich Oligonucleotides Correlates with Protein Binding

Bates

Kahlon

Thomas

et al. 1999

Journal of Biological Chemistry

356

363

View full text Add to dashboard Cite

Oligonucleotides have been extensively studied as antisense or antigene agents that can potentially modulate the expression of specific genes. These strategies rely on sequence-specific hybridization of the oligonucleotide to mRNA or genomic DNA. Recently, it has become clear that oligonucleotides often have biological activities that cannot be attributed to their sequence-specific interactions with nucleic acids. Here we describe a series of guanosine-rich phosphodiester oligodeoxynucleotides that strongly inhibit proliferation in a number of human tumor cell lines. The presence of G-quartets in the active oligonucleotides is demonstrated using an UV melting technique. We show that G-rich oligonucleotides bind to a specific cellular protein and that the biological activity of the oligonucleotides correlates with binding to this protein. The G-rich oligonucleotidebinding protein was detected in both nuclear and cytoplasmic extracts and in proteins derived from the plasma membrane of cells. We present strong evidence that this protein is nucleolin, a multifunctional phosphoprotein whose levels are related to the rate of cell proliferation. Our results indicate that binding of G-rich oligonucleotides to nucleolin may be responsible for their non-sequence-specific effects. Furthermore, these oligonucleotides represent a new class of potentially therapeutic agents with a novel mechanism of action.

show abstract

“…Considering the polyanionic features of ASOs, the difficulty of these compounds to cross the plasmamembrane passively can be understood. The uptake of ASOs occurs through active transport, which, in turn, depends on the temperature (Yakubov et al, 1989), the structure and the concentration of oligos (Vlassov et al, 1994). Several approaches have been explored to enhance cellular uptake and permeability of ASOs in order to increase their therapeutic efficacy, including conjugation to polycations, targeting cell surface receptors, coupling to cholesterol, encapsulation into lipid particles such as microspheres and liposomes.…”

Section: Antisense Therapy: Rationale and Limitationsmentioning

confidence: 99%