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 Handhold-Mediated Strand Displacement: A Nucleic Acid Based Mechanism for Generating Far-from-Equilibrium Assemblies through Templated Reactions

ACS Nano ◽  
2021 ◽  
Vol 15 (2) ◽  
pp. 3272-3283
Author(s):  
Javier Cabello-Garcia ◽  
Wooli Bae ◽  
Guy-Bart V. Stan ◽  
Thomas E. Ouldridge
Keyword(s):  
Nucleic Acid ◽  
Strand Displacement ◽  
Far From Equilibrium

scholarly journals Conditional guide RNA through two intermediate hairpins for programmable CRISPR/Cas9 function: building regulatory connections between endogenous RNA expressions

2020 ◽  
Vol 48 (20) ◽  
pp. 11773-11784
Author(s):  
Jiao Lin ◽  
Yan Liu ◽  
Peidong Lai ◽  
Huixia Ye ◽  
Liang Xu
Keyword(s):  
Nucleic Acid ◽  
Genetic Regulation ◽  
Binding Activity ◽  
Strand Displacement ◽  
Genetic Circuits ◽  
Reporter System ◽  
Gene Expressions ◽  
Guide Rna ◽  
Naturally Occurring ◽  
Novel Approach

Abstract A variety of nanodevices developed for nucleic acid computation provide great opportunities to construct versatile synthetic circuits for manipulation of gene expressions. In our study, by employing a two-hairpin mediated nucleic acid strand displacement as a processing joint for conditional guide RNA, we aim to build artificial connections between naturally occurring RNA expressions through programmable CRISPR/Cas9 function. This two-hairpin joint possesses a sequence-switching machinery, in which a random trigger strand can be processed to release an unconstrained sequence-independent strand and consequently activate the self-inhibitory guide RNA for conditional gene regulation. This intermediate processor was characterized by the fluorescence reporter system and applied for regulation of the CRISPR/Cas9 binding activity. Using plasmids to generate this sequence-switching machinery in situ, we achieved the autonomous genetic regulation of endogenous RNA expressions controlled by other unrelated endogenous RNAs in both E. coli and human cells. Unlike previously reported strand-displacement genetic circuits, this advanced nucleic acid nanomachine provides a novel approach that can establish regulatory connections between naturally occurring endogenous RNAs. In addition to CRISPR systems, we anticipate this two-hairpin machine can serve as a general processing joint for wide applications in the development of other RNA-based genetic circuits.

scholarly journals Protein-Based Systems for Translational Regulation of Synthetic mRNAs in Mammalian Cells

Life ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1192
Author(s):  
Hideyuki Nakanishi
Keyword(s):  
Adverse Effects ◽  
Mammalian Cells ◽  
Translational Regulation ◽  
Insertional Mutagenesis ◽  
In Vitro Transcription ◽  
Preclinical Studies ◽  
High Efficacy ◽  
Gene Circuits ◽  
Synthetic Mrna

Synthetic mRNAs, which are produced by in vitro transcription, have been recently attracting attention because they can express any transgenes without the risk of insertional mutagenesis. Although current synthetic mRNA medicine is not designed for spatiotemporal or cell-selective regulation, many preclinical studies have developed the systems for the translational regulation of synthetic mRNAs. Such translational regulation systems will cope with high efficacy and low adverse effects by producing the appropriate amount of therapeutic proteins, depending on the context. Protein-based regulation is one of the most promising approaches for the translational regulation of synthetic mRNAs. As synthetic mRNAs can encode not only output proteins but also regulator proteins, all components of protein-based regulation systems can be delivered as synthetic mRNAs. In addition, in the protein-based regulation systems, the output protein can be utilized as the input for the subsequent regulation to construct multi-layered gene circuits, which enable complex and sophisticated regulation. In this review, I introduce what types of proteins have been used for translational regulation, how to combine them, and how to design effective gene circuits.

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.

scholarly journals A nucleic acid strand displacement system for the multiplexed detection of tuberculosis-specific mRNA using quantum dots

Nanoscale ◽  
2016 ◽  
Vol 8 (19) ◽  
pp. 10087-10095 ◽  
Author(s):  
H. D. Gliddon ◽  
P. D. Howes ◽  
M. Kaforou ◽  
M. Levin ◽  
M. M. Stevens
Keyword(s):  
Quantum Dots ◽  
Nucleic Acid ◽  
Strand Displacement ◽  
Multiplexed Detection ◽  
Mrna Detection ◽  
Dna Strand Displacement ◽  
Displacement System ◽  
Dna Strand ◽  
Specific Mrna

On the development of a novel multiplexed assay for Tuberculosis-specific mRNA detection using DNA strand displacement and quantum dots.

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

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 Mismatches Improve the Performance of Strand-Displacement Nucleic Acid Circuits

2014 ◽  
Vol 126 (7) ◽  
pp. 1876-1879 ◽  
Author(s):  
Yu Sherry Jiang ◽  
Sanchita Bhadra ◽  
Bingling Li ◽  
Andrew D. Ellington
Keyword(s):  
Nucleic Acid ◽  
Strand Displacement
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