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Contract number
14.W03.31.0007
Time span of the project
2017-2019
Head of the laboratory

As of 30.01.2020

9
Number of staff members
12
scientific publications
General information

Name of the project: Study of mechanisms of RNA interference and protection of genomes in bacteria and eukaryotes.

Strategy for Scientific and Technological Development Priority Level: в


Goals and objectives

Research directions: Biogenesis of non-coding RNA in eukaryotes, RNA interference in bacteria, genome editing using non-coding RNA and Argonaute proteins

Project objective: Study of natural function of RNA interference systems in bacteria and eukaryotes, as well as development of new tools for gene and electronic engineering based on these systems


The practical value of the study

  • We have produced transgenic lines pf drosophila with expression of RNA processing proteins containing affinity tag and refined a methodology of profiling of RNA-binding proteins for analysis of RNA-protein interactions.
  • We have studied a new genome locus, responsible for production of piРНКof male hormonal cells of drosophila. It has been shown that this locus facilitates reproductive isolation between closely related types of drosophila.
  • Our have produced human cell lines with integration of barcoded transgenes into genome. Those cell lines allows to research correlation between structure of chromatine and expression of reporter genes.
  • We have conducted bioinformatical analysis of bacterial Argonaute proteins and factor associated with them, and performed their classification.
  • Our Laboratory has created a collection of natural bacteria containing Argonaute proteins of various classes and cloned genes of new Argonaute proteins.
  • It has been shown that bacterial Argonaute proteins are DNA-dependent DNA nucleases and are able to perform cutting of DNA targets in vitro. We have researched factors determining efficiency of processing of DNA by these proteins.
  • We have conducted analysis of libraries of DNA associated with Argonaute proteins in cells of bacteria and studied factors determining specificity of processing of DNA by these proteins.
  • Our researchers have developed experimental approaches to analysis of specificity of Argonaute proteins in vivo and study of their role protecting cells of bacteria from mobile genetic elements and bacteriophages.
  • We have conducted expression of Argonaute proteins in eukaryotic cells and produced optimized variations of Argonautes for their usage in gene editing.

IEducation and career development:

  • We have provided training to employees and postgraduates of the Laboratory in new methods of research by organizing academic trips to the California Institute of Technology (USA).
  • 5 postgraduate students have been hired by the hosting foundation.

Collaborations:

  • California Institute of Technology (USA): joint research, internships of postgraduates and young scientists
  • Memorial Sloan-Kettering Cancer Center (Dinshaw Patel) (USA), Icahn School of Medicine at Mount Sinai (Ravi Sachidanandam) (USA), Laboratory of Cell Division of the Institute of Moledulcar and Cell Biology of the SIberian Department of the Russian Academy of Sciences (Russia): joint research

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Lisitskaya L, Aravin A.A., Kulbachinskiy A.
DNA interference and beyond: structure and functions of prokaryotic Argonaute proteins. Nature Communications 9(1): 5165 (2018).
Ryazansky S., Kulbachinskiy A., Aravin A.A.
The Expanded Universe of Prokaryotic Argonaute Proteins. MBio 9(6): e01935-18 (2018).
Pupov A., Ignatov A., Agapov A., Kulbachinskiy A.
Distinct effects of DNA lesions on RNA synthesis by Escherichia coli RNA polymerase. Biochemical and Biophysical Research Communications 510(1): 122-127 (2019).
Kotov A.A., Adashev V.E., Godneeva B.K., Ninova M., Shatskikh A.S., Bazylev S.S., Aravin A.A., Olenina L.V
piRNA silencing contributes to interspecies hybrid sterility and reproductive isolation in Drosophila melanogaster. Nucleic Acids Research, gkz130 (2019).
Mindlin S., Kudinova A., Mardanov A., Petrova M.
Adaptive dif modules in permafrost strains of Acinetobacter lwoffii and their distribution and abundance among present day Acinetobacter strains. Frontiers in Microbiology, 29 March 2019.
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