Group of Microbial Genoms Editing

Anna V. Goncharenko
Head of the Group
Ph.D. (Biology)
INBI, build.3, room. 10
Telefone+7 (495) 660-34-30 ext. 117
E-Mail pylaevanna@gmail.com

Key words:
genome editing, CRISPR / Cas systems, homologous recombination, NHEJ DNA repair, mycobacteria, latent tuberculosis, persistence, toxin-antitoxin loci, BCG, live recombinant vaccines.

Research areas:

  • Applying CRISPR / Cas for editing mycobacterial genomes
  • CRISPR / Cas interference
  • Mycobacterium toxin-anti-toxin systems as regulators of persistence and transition of mycobacteria to the state of metabolic dormancy
  • Creation of recombinant BCG strains with improved protective properties

Basic research methods:
Molecular cloning, generation of gene knockouts by allelic exchange, CRISPR editing of bacterial genomes, creation of strains overexpressing target genes, and constructs for the heterologous expression of recombinant proteins, site-directed mutagenesis, Northern and Western blotting, Primer extension analysis, quantitative real-time PCR, microbiological methods for the study of recombinant bacterial strains.

Origins:
The group was created in March 2019 during the reorganization of the laboratory of Biochemistry of Stresses in Microorganisms. Now it is the part of the Department of Medical Microbiology in Research Center of Biotechnology RAS.

 

Relevant publications related to the group research directions:

  1. Akhova A., Nesterova L., Shumkov M., Tkachenko A. Cadaverine biosynthesis contributes to decreased Escherichia coli susceptibility to antibiotics // Research in Microbiology. 2021; 172(7-8): 103881. DOI: 10.1016/j.resmic.2021.103881.
  2. Karpov D. S., Goncharenko A. V., Usachev E. V., Vasina D. V., Divisenko E. V., Chalenko Y. M., Pochtovyi A. A., Ovchinnikov R. S., Makarov V. V., Yudin S. M., Tkachuk A. P., Gushchin V. A. A Strategy for the Rapid Development of a Safe Vibrio cholerae Candidate Vaccine Strain // International journal of molecular sciences. 2021; 22(21): 11657. DOI: 10.3390/ijms222111657.
  3. Zamakhaev M., Tsyganov I., Nesterova L., Akhova A., Grigorov A., Bespyatykh J., Azhikina T., Tkachenko A., Shumkov M. Mycolicibacterium smegmatis possesses operational agmatinase but contains no detectable polyamines // International Journal of Mycobacteriology. 2020; 9(2): 138-143. DOI: 10.4103/ijmy.ijmy_48_20.
  4. Margarita Shleeva, Tatyana Kondratieva, Galina Demina, Elvira Rubakova, Anna Goncharenko, Alexander Apt, Arseny Kaprelyants. Overexpression of Adenylyl Cyclase Encoded by the Mycobacterium tuberculosis Rv2212 Gene Confers Improved Fitness, Accelerated Recovery from Dormancy and Enhanced Virulence in Mice, FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, 2017, v 7
  5. Alexander G. Tkachenko, Natalya M. Kashevarova, Elena A. Tyuleneva, Mikhail S. Shumkov; Stationary-phase genes upregulated by polyamines are responsible for the formation of Escherichia coli persister cells tolerant to netilmicin. // FEMS Microbiol Lett, 2017
  6. Soluble expression and easy purification of human bone morphogenetic protein-2, Goncharenko A., Prilipov A., Sadykova G., Moisenovich M., Kirpichnikov M. The FEBS Journal, 284 (Suppl. 1), p. 360, 2017
  7. M. Zamakhaev, J. Bespyatykh, A. Goncharenko, M. Shumkov, “How to turn it off? Role of VapC toxin in mycobacteria dormant state formation”, The FEBS Journal, vol. 284 (Suppl. 1), p. 360, 2017
  8. Toxin-antitoxin vapBC locus participates in formation of the dormant state in Mycobacterium smegmatis. Demidenok OI, Kaprelyants AS, Goncharenko AV. FEMS Microbiol Lett. 2014 Jan 13
  9. Cyclic amp-dependent resuscitation of dormant mycobacteria by exogenous free Fatty acids. Shleeva M, Goncharenko A, Kudykina Y, Young D, Young M, Kaprelyants A. PLoS One. 2013 Dec 23;8(12)
  10. Anuchin AM, Goncharenko AV, Demina GR, Mulyukin AL, Ostrovsky DN, Kaprelyants AS. The role of histone-like protein, Hlp, in Mycobacterium smegmatis dormancy. FEMS Microbiol Lett. 2010 Jul;308(2):101-7
  11. Koltunov V, Greenblatt CL, Goncharenko AV, Demina GR, Klein BY, Young M, Kaprelyants AS. Structural changes and cellular localization of resuscitation-promoting factor in environmental isolates of Micrococcus luteus. Microb Ecol. 2010 Feb;59(2):296-310