Nucleic Acid Electrotransfer in Mammalian Cells: Mechanistic Description

2017 ◽  
pp. 323-336 ◽  
Author(s):  
Muriel Golzio ◽  
Marie-Pierre Rols
Keyword(s):  
Nucleic Acid ◽  
Mammalian Cells

scholarly journals Nucleic Acid Strand Displacement with Synthetic mRNA Inputs in Living Mammalian Cells

2018 ◽  
Vol 7 (12) ◽  
pp. 2737-2741 ◽  
Author(s):  
Gourab Chatterjee ◽  
Yuan-Jyue Chen ◽  
Georg Seelig
Keyword(s):  
Nucleic Acid ◽  
Mammalian Cells ◽  
Strand Displacement ◽  
Synthetic Mrna

scholarly journals 8-AZAGUANINE RESISTANCE IN MAMMALIAN CELLS I. HYPOXANTHINE-GUANINE PHOSPHORIBOSYLTRANSFERASE

Genetics ◽  
1972 ◽  
Vol 72 (2) ◽  
pp. 239-252 ◽  
Author(s):  
F D Gillin ◽  
D J Roufa ◽  
A L Beaudet ◽  
C T Caskey
Keyword(s):  
Nucleic Acid ◽  
Mammalian Cells ◽  
Chinese Hamster ◽  
Acid Synthesis ◽  
Ethyl Methanesulfonate ◽  
Nucleic Acid Synthesis ◽  
Wild Type ◽  
Chinese Hamster Cells ◽  
Hypoxanthine Guanine Phosphoribosyltransferase

ABSTRACT Chinese hamster cells were treated with ethyl methanesulfonate or N-methyl-N'-nitro-N-nitrosoguanidine, and mutants resistant to 8-azaguanine were selected and characterized. Hypoxanthine-guanine phosphoribosyltransferase activity of sixteen mutants is extremely negative, making them suitable for reversion to HGPRTase+. Ten of the extremely negative mutants revert at a frequency higher than 10-7 suggesting their point mutational character. The remaining mutants have demonstrable HGPRTase activity and are not useful for reversion analysis. Five of these mutants have < 2% HGPRTase and are presumably also HGPRTase point mutants. The remaining 14 mutants utilize exogenous hypoxanthine for nucleic acid synthesis poorly, and possess 20-150% of wild-type HGPRTase activity in in vitro. Their mechanism of 8-azaguanine resistance is not yet defined.

Metabolic Co-Operation Between Biochemically Marked Mammalian Cells in Tissue Culture

1969 ◽  
Vol 4 (2) ◽  
pp. 353-367
Author(s):  
H. SUBAK-SHARPE ◽  
R. R. BÜRK ◽  
J. D. PITTS
Keyword(s):  
Tissue Culture ◽  
Nucleic Acid ◽  
Mixed Culture ◽  
Molecular Basis ◽  
Mammalian Cells ◽  
Genetic Variant ◽  
Cell Contact ◽  
Metabolic Function ◽  
Indirect Contact ◽  
Fibroblast Line

Cells of a genetic variant of the hamster fibroblast line BHK 21 which lack inosinic pyrophosphorylase activity (IPP- cells) and therefore cannot normally incorporate [3H]hypoxanthine were grown in mixed culture with cells of BHK 21 sublines which have inosinic pyrophosphorylase activity (IPP+ cells). If not in contact with IPP+ cells, IPP- cells do not incorporate added [3H]hypoxanthine into nucleic acid. IPP+ cells always do incorporate [3H]hypoxanthine and IPP- cells when in direct or indirect contact with IPP+ cells also incorporate the isotope. Cell to cell contact appears to be essential for this gain of a metabolic function by IPP- cells. The possible molecular basis and general implications of the phenomenon are discussed.

RNA interference by 2′,5′-linked nucleic acid duplexes in mammalian cells

2006 ◽  
Vol 16 (12) ◽  
pp. 3238-3240 ◽  
Author(s):  
Thazha P. Prakash ◽  
Bryan Kraynack ◽  
Brenda F. Baker ◽  
Eric E. Swayze ◽  
Balkrishen Bhat
Keyword(s):  
Rna Interference ◽  
Nucleic Acid ◽  
Mammalian Cells

scholarly journals VLPs Derived from the CCMV Plant Virus Can Directly Transfect and Deliver Heterologous Genes for Translation into Mammalian Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
María V. Villagrana-Escareño ◽  
Elizabeth Reynaga-Hernández ◽  
Othir G. Galicia-Cruz ◽  
Ana L. Durán-Meza ◽  
Viridiana De la Cruz-González ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Mammalian Cells ◽  
Plant Virus ◽  
Fluorescent Protein ◽  
Genetic Material ◽  
Eukaryotic Cell ◽  
Plant Viruses ◽  
Virus Capsids ◽  
First Time ◽  
Drug Nanocarriers

Virus-like particles (VLPs) are being used for therapeutic developments such as vaccines and drug nanocarriers. Among these, plant virus capsids are gaining interest for the formation of VLPs because they can be safely handled and are noncytotoxic. A paradigm in virology, however, is that plant viruses cannot transfect and deliver directly their genetic material or other cargos into mammalian cells. In this work, we prepared VLPs with the CCMV capsid and the mRNA-EGFP as a cargo and reporter gene. We show, for the first time, that these plant virus-based VLPs are capable of directly transfecting different eukaryotic cell lines, without the aid of any transfecting adjuvant, and delivering their nucleic acid for translation as observed by the presence of fluorescent protein. Our results show that the CCMV capsid is a good noncytotoxic container for genome delivery into mammalian cells.

scholarly journals Polyamines and Their Role in Virus Infection

2017 ◽  
Vol 81 (4) ◽  
Author(s):  
Bryan C. Mounce ◽  
Michelle E. Olsen ◽  
Marco Vignuzzi ◽  
John H. Connor
Keyword(s):  
Protein Synthesis ◽  
Nucleic Acid ◽  
Mammalian Cells ◽  
Cellular Proliferation ◽  
Polyamine Synthesis ◽  
Host Interactions ◽  
Dna And Rna ◽  
Rna Polymerization ◽  
Nucleic Acid Conformation

SUMMARY Polyamines are small, abundant, aliphatic molecules present in all mammalian cells. Within the context of the cell, they play a myriad of roles, from modulating nucleic acid conformation to promoting cellular proliferation and signaling. In addition, polyamines have emerged as important molecules in virus-host interactions. Many viruses have been shown to require polyamines for one or more aspects of their replication cycle, including DNA and RNA polymerization, nucleic acid packaging, and protein synthesis. Understanding the role of polyamines has become easier with the application of small-molecule inhibitors of polyamine synthesis and the use of interferon-induced regulators of polyamines. Here we review the diverse mechanisms in which viruses require polyamines and investigate blocking polyamine synthesis as a potential broad-spectrum antiviral approach.

scholarly journals Assembly Properties of Human Immunodeficiency Virus Type 1 Gag-Leucine Zipper Chimeras: Implications for Retrovirus Assembly

2008 ◽  
Vol 83 (5) ◽  
pp. 2216-2225 ◽  
Author(s):  
Rachael M. Crist ◽  
Siddhartha A. K. Datta ◽  
Andrew G. Stephen ◽  
Ferri Soheilian ◽  
Jane Mirro ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Nucleic Acid ◽  
Human Immunodeficiency Virus Type ◽  
Mammalian Cells ◽  
Leucine Zipper ◽  
Chimeric Proteins ◽  
Wild Type ◽  
Particle Assembly ◽  
Immunodeficiency Virus

ABSTRACT Expression of the retroviral Gag protein leads to formation of virus-like particles in mammalian cells. In vitro and in vivo experiments show that nucleic acid is also required for particle assembly. However, several studies have demonstrated that chimeric proteins in which the nucleocapsid domain of Gag is replaced by a leucine zipper motif can also assemble efficiently in mammalian cells. We have now analyzed assembly by chimeric proteins in which nucleocapsid of human immunodeficiency virus type 1 (HIV-1) Gag is replaced by either a dimerizing or a trimerizing zipper. Both proteins assemble well in human 293T cells; the released particles lack detectable RNA. The proteins can coassemble into particles together with full-length, wild-type Gag. We purified these proteins from bacterial lysates. These recombinant “Gag-Zipper” proteins are oligomeric in solution and do not assemble unless cofactors are added; either nucleic acid or inositol phosphates (IPs) can promote particle assembly. When mixed with one equivalent of IPs (which do not support assembly of wild-type Gag), the “dimerizing” Gag-Zipper protein misassembles into very small particles, while the “trimerizing” protein assembles correctly. However, addition of both IPs and nucleic acid leads to correct assembly of all three proteins; the “dimerizing” Gag-Zipper protein also assembles correctly if inositol hexakisphosphate is supplemented with other polyanions. We suggest that correct assembly requires both oligomeric association at the C terminus of Gag and neutralization of positive charges near its N terminus.

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