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39 results on '"Gurskaya, Nadya G."'

10. hTERT-Driven Immortalization of RDEB Fibroblast and Keratinocyte Cell Lines Followed by Cre-Mediated Transgene Elimination

Author

Evtushenko, Nadezhda A., primary, Beilin, Arkadii K., additional, Dashinimaev, Erdem B., additional, Ziganshin, Rustam H., additional, Kosykh, Anastasiya V., additional, Perfilov, Maxim M., additional, Rippa, Alexandra L., additional, Alpeeva, Elena V., additional, Vasiliev, Andrey V., additional, Vorotelyak, Ekaterina A., additional, and Gurskaya, Nadya G., additional

Published
2021
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11. Signatures of Dermal Fibroblasts from RDEB Pediatric Patients

Author

Beilin, Arkadii K., primary, Evtushenko, Nadezhda A., additional, Lukyanov, Daniil K., additional, Murashkin, Nikolay N., additional, Ambarchian, Eduard T., additional, Pushkov, Alexander A., additional, Savostyanov, Kirill V., additional, Fisenko, Andrey P., additional, Rogovaya, Olga S., additional, Vasiliev, Andrey V., additional, Vorotelyak, Ekaterina A., additional, and Gurskaya, Nadya G., additional

Published
2021
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13. Genetically Encoded Fluorescent Sensor for Poly-ADP-Ribose

Author

Serebrovskaya, Ekaterina O., primary, Podvalnaya, Nadezda M., additional, Dudenkova, Varvara V., additional, Efremova, Anna S., additional, Gurskaya, Nadya G., additional, Gorbachev, Dmitry A., additional, Luzhin, Artem V., additional, Kantidze, Omar L., additional, Zagaynova, Elena V., additional, Shram, Stanislav I., additional, and Lukyanov, Konstantin A., additional

Published
2020
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17. Functioning of Fluorescent Proteins in Aggregates in Anthozoa Species and in Recombinant Artificial Models.

Author

Povarova, Natalia V., Petri, Natalia D., Blokhina, Anna E., Bogdanov, Alexey M., Gurskaya, Nadya G., and Lukyanov, Konstantin A.

Subjects
FLUORESCENT proteins, ANTHOZOA, CONFOCAL microscopy, PROTEINS, INTRACELLULAR membranes
Abstract

Despite great advances in practical applications of fluorescent proteins (FPs), their natural function is poorly understood. FPs display complex spatio-temporal expression patterns in living Anthozoa coral polyps. Here we applied confocal microscopy, specifically, the fluorescence recovery after photobleaching (FRAP) technique to analyze intracellular localization and mobility of endogenous FPs in live tissues. We observed three distinct types of protein distributions in living tissues. One type of distribution, characteristic for Anemonia, Discosoma and Zoanthus, is free, highly mobile cytoplasmic localization. Another pattern is seen in FPs localized to numerous intracellular vesicles, observed in Clavularia. The third most intriguing type of intracellular localization is with respect to the spindle-shaped aggregates and lozenge crystals several micrometers in size observed in Zoanthus samples. No protein mobility within those structures was detected by FRAP. This finding encouraged us to develop artificial aggregating FPs. We constructed "trio-FPs" consisting of three tandem copies of tetrameric FPs and demonstrated that they form multiple bright foci upon expression in mammalian cells. High brightness of the aggregates is advantageous for early detection of weak promoter activities. Simultaneously, larger aggregates can induce significant cytostatic and cytotoxic effects and thus such tags are not suitable for long-term and high-level expression. [ABSTRACT FROM AUTHOR]

Published
2017
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20. Structural Basis for Phototoxicity of the Genetically Encoded Photosensitizer KillerRed

Author

Pletnev, Sergei, primary, Gurskaya, Nadya G., additional, Pletneva, Nadya V., additional, Lukyanov, Konstantin A., additional, Chudakov, Dmitri M., additional, Martynov, Vladimir I., additional, Popov, Vladimir O., additional, Kovalchuk, Mikhail V., additional, Wlodawer, Alexander, additional, Dauter, Zbigniew, additional, and Pletnev, Vladimir, additional

Published
2009
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31. Natural Animal Coloration Can Be Determined by a Nonfluorescent Green Fluorescent Protein Homolog

Author

Lukyanov, Konstantin A., primary, Fradkov, Arkady F., additional, Gurskaya, Nadya G., additional, Matz, Mikhail V., additional, Labas, Yulii A., additional, Savitsky, Aleksandr P., additional, Markelov, Mikhail L., additional, Zaraisky, Andrey G., additional, Zhao, Xiaoning, additional, Fang, Yu, additional, Tan, Wenyan, additional, and Lukyanov, Sergey A., additional

Published
2000
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32. Color transitions in coral's fluorescent proteins by site-directed mutagenesis

Author

Lukyanov Sergey A, Yanushevich Yurii G, Savitsky Alexander P, Gurskaya Nadya G, and Lukyanov Konstantin A

Subjects
Green Fluorescent Proteins, Molecular Sequence Data, lcsh:Animal biochemistry, Color, Anthozoa, Fluorescence, lcsh:Biochemistry, Luminescent Proteins, Spectrometry, Fluorescence, Amino Acid Substitution, Microscopy, Fluorescence, Escherichia coli, Mutagenesis, Site-Directed, lcsh:QD415-436, Amino Acid Sequence, lcsh:QP501-801, Sequence Alignment, Research Article
Abstract

Background Green Fluorescent Protein (GFP) cloned from jellyfish Aequorea victoria and its homologs from corals Anthozoa have a great practical significance as in vivo markers of gene expression. Also, they are an interesting puzzle of protein science due to an unusual mechanism of chromophore formation and diversity of fluorescent colors. Fluorescent proteins can be subdivided into cyan (~ 485 nm), green (~ 505 nm), yellow (~ 540 nm), and red (>580 nm) emitters. Results Here we applied site-directed mutagenesis in order to investigate the structural background of color variety and possibility of shifting between different types of fluorescence. First, a blue-shifted mutant of cyan amFP486 was generated. Second, it was established that cyan and green emitters can be modified so as to produce an intermediate spectrum of fluorescence. Third, the relationship between green and yellow fluorescence was inspected on closely homologous green zFP506 and yellow zFP538 proteins. The following transitions of colors were performed: yellow to green; yellow to dual color (green and yellow); and green to yellow. Fourth, we generated a mutant of cyan emitter dsFP483 that demonstrated dual color (cyan and red) fluorescence. Conclusions Several amino acid substitutions were found to strongly affect fluorescence maxima. Some positions primarily found by sequence comparison were proved to be crucial for fluorescence of particular color. These results are the first step towards predicting the color of natural GFP-like proteins corresponding to newly identified cDNAs from corals.

Published
2001

33. Equalizing cDNA Subtraction Based on Selective Suppression of Polymerase Chain Reaction: Cloning of Jurkat Cell Transcripts Induced by Phytohemaglutinin and Phorbol 12-Myristate 13-Acetate

Author

Gurskaya, Nadya G., primary, Diatchenko, Luda, additional, Chenchik, Alex, additional, Siebert, Paul D., additional, Khaspekov, George L., additional, Lukyanov, Konstantin A., additional, Vagner, Laura L., additional, Ermolaeva, Olga D., additional, Lukyanov, Sergey A., additional, and Sverdlov, Eugene D., additional

Published
1996
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34. A strategy for the generation of non-aggregating mutants of Anthozoa fluorescent proteins

Author

Yanushevich, Yurii G., Staroverov, Dmitry B., Savitsky, Alexander P., Fradkov, Arkady F., Gurskaya, Nadya G., Bulina, Maria E., Lukyanov, Konstantin A., and Lukyanov, Sergey A.

Subjects
GREEN fluorescent protein, MUTAGENESIS
Abstract

Recently, we cloned several fluorescent proteins of different colors homologous to Aequorea victoria green fluorescent protein, which have great biotechnological potential as in vivo markers of gene expression. However, later investigations revealed severe drawbacks in the use of novel fluorescent proteins (FPs), in particular, the formation of tetramers (tetramerization) and high molecular weight aggregates (aggregation). In this report, we employ a mutagenic approach to resolve the problem of aggregation. The elimination of basic residues located near the N-termini of FPs results in the generation of non-aggregating versions of several FPs, specifically, drFP583 (DsRed), DsRed-Timer, ds/drFP616, zFP506, zFP538, amFP486, and asFP595. [Copyright &y& Elsevier]

Published
2002
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35. GFP-like chromoproteins as a source of far-red fluorescent proteins11The accession numbers of genes described in this article are: cgCP (AF363775), hcCP (AF363776), cpCP (AF383155), gtCP (AF383156)

Author

Gurskaya, Nadya G., Fradkov, Arkady F., Terskikh, Alexey, Matz, Mikhail V., Labas, Yulii A., Martynov, Vladimir I., Yanushevich, Yurii G., Lukyanov, Konstantin A., and Lukyanov, Sergey A.

Subjects
Multi-color labeling, drFP583, DsRed, Green fluorescent protein, Anthozoa
Abstract

We have employed a new approach to generate novel fluorescent proteins (FPs) from red absorbing chromoproteins. An identical single amino acid substitution converted novel chromoproteins from the species Anthozoa (Heteractis crispa, Condylactis gigantea, and Goniopora tenuidens) into far-red FPs (emission λmax=615–640 nm). Moreover, coupled site-directed and random mutagenesis of the chromoprotein from H. crispa resulted in a unique far-red FP (HcRed) that exhibited bright emission at 645 nm. A clear red shift in fluorescence of HcRed, compared to drFP583 (by more than 60 nm), makes it an ideal additional color for multi-color labeling. Importantly, HcRed is excitable by 600 nm dye laser, thus promoting new detection channels for multi-color flow cytometry applications. In addition, we generated a dimeric mutant with similar maturation and spectral properties to tetrameric HcRed.

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36. Method for quantitative analysis of nonsense-mediated mRNA decay at the single cell level.

Author

Kudryavtseva, Elena I., Lukyanov, Konstantin A., Pereverzev, Anton P., Gurskaya, Nadya G., Ermakova, Galina V., Kotlobay, Alexey A., Zaraisky, Andrey G., Markina, Nadezhda M., and Lukyanov, Sergey A.

Subjects
MESSENGER RNA, CELL lines, CYTOMETRY, FLUORESCENCE microscopy, FLUORESCENT proteins, GENETIC regulation, XENOPUS
Abstract

Nonsense-mediated mRNA decay (NMD) is a ubiquitous mechanism of degradation of transcripts with a premature termination codon. NMD eliminates aberrant mRNA species derived from sources of genetic variation such as gene mutations, alternative splicing and DNA rearrangements in immune cells. In addition, recent data suggest that NMD is an important mechanism of global gene expression regulation. Here, we describe new reporters to quantify NMD activity at the single cell level using fluorescent proteins of two colors: green TagGFP2 and far-red Katushka. TagGFP2 was encoded by mRNA targeted to either the splicing-dependent or the long 3'UTR-dependent NMD pathway. Katushka was used as an expression level control. Comparison of the fluorescence intensities of cells expressing these reporters and cells expressing TagGFP2 and Katushka from corresponding control NMD-independent vectors allowed for the assessment of NMD activity at the single cell level using fluorescence microscopy and flow cytometry. The proposed reporter system was successfully tested in several mammalian cell lines and in transgenic Xenopus embryos. [ABSTRACT FROM AUTHOR]

Published
2015
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39. Generation of Cell Lines Stably Expressing a Fluorescent Reporter of Nonsense-Mediated mRNA Decay Activity.

Author

Markina NM, Pereverzev AP, Staroverov DB, Lukyanov KA, and Gurskaya NG

Subjects
Animals, Cell Separation instrumentation, Cell Separation methods, Flow Cytometry instrumentation, Flow Cytometry methods, Green Fluorescent Proteins genetics, Humans, Mice, Microscopy, Fluorescence methods, RNA Splicing genetics, RNA, Messenger metabolism, Transfection methods, Transgenes genetics, Cell Line metabolism, Genes, Reporter genetics, Genetic Engineering methods, Green Fluorescent Proteins metabolism, Nonsense Mediated mRNA Decay genetics
Abstract

Nonsense-mediated mRNA decay (NMD) is a mechanism of mRNA surveillance ubiquitous among eukaryotes. Importantly, NMD not only removes aberrant transcripts with premature stop codons, but also regulates expression of many normal genes. A recently introduced dual-color fluorescent protein-based reporter enables analysis of NMD activity in live cells. In this chapter we describe the method to generate stable transgenic cell lines expressing the splicing-dependent NMD reporter using consecutive steps of lentivirus transduction and Tol2 transposition.

Published
2018
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