`Yuri Voziyanov. Gene editing. Genome Engineering. Flp - FRT, Cre - loxP, TD - TDRT, R - R1RT site-specific DNA recombination
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The Jackson Laboratory



Research Tools


Feng Li
Genome engineering
Evolution of variants of Flp recombinase

Riddhi Shah
Genome engineering
Evolution of variants of Flp and Flp-like recombinases

Yue Li
Genome engineering

Eugenia Voziyanova
Evolution of variants of Flp recombinase
Cell replacement

Yuri Voziyanov
Professor of Biology

Marvin T. Green, Sr.
Endowed Professor
in Pre-medicine

Louisiana Tech University
School of Biological Sciences / Institute for Micromanufacturing
CTH 121
1 Adams Blvd.
Ruston, LA 71272

318.257.2694
voziyan@latech.edu

1991 - BS/MS, Kiev State University, Kiev, Ukraine
1996 - Ph.D., Institute of Molecular Biology and Genetics, Kiev, Ukraine
1997 - 2003 - Postdoctoral training, University of Texas at Austin and European Molecular Biology Laboratory, Heidelberg, Germany



RESEARCH


There are three directions of our current research: gene editing and genome engineering, protein engineering, and aging.

Gene editing and genome engineering

Our main research direction is gene editing aimed to repair genetic defects in individuals with genetic diseases. Currently, we are working on correcting mutations that cause sickle cell anemia disease. In our experiments we use several gene editing/genome engineering tools: CRISPR/Cas9 and TALENs systems along with the target-specific variants of site-specific DNA recombinases: Flp, Cre, and related recombinases. For our genome engineering experiments, we design non-viral vectors with different functional elements, including S/MAR and insulator elements. We also experiment with in vitro transcribed mRNA, shRNA, transposase-mediated integrating vectors, and minicircles.

Protein engineering

Our second research direction is protein engineering. We evolve site-specific DNA recombinases of tyrosine type with new target specificity and analyze how individual amino acid substitutions influence the specificity switch. In this research we use popular site-specific recombinases Flp, Cre, as well as newly introduced TD and R. The tyrosine site-specific recombinases can mediate a full variety of DNA rearrangement reaction: integration and excision, inversion, translocation, and cassette replacement. These enzymes are particularly widespread in bacteria: over 1000 tyrosine site-specific recombinases are identified. All these enzymes have similar 3D fold but remarkably different target specificity. These properties make tyrosine recombinases an attractive model for changing target specificity of an enzyme and deciphering the functional role of their amino acid residues in target recognition. Moreover, the recombinase variants with modified target specificity could become useful gene editing and genome engineering tools.

Aging

Our third emerging research direction is aging. In this reseach we want to understand how to get rid of the harmful modifications that cells acquire during the process aging. As a model organism, we use budding yeast as they are able to reset their lifespan during spore formation.



[Publications (PubMed database)]

Selected publications


  • Voziyanova E., Anderson R.P., Shah R., Li F., Voziyanov Y.
    Efficient genome manipulation by variants of site-specific recombinases R and TD.
    J. Mol. Biol., 2016

  • Shah R., Li F., Voziyanova E., Voziyanov Y.
    Target-specific variants of Flp recombinase mediate genome engineering reactions in mammalian cells.
    FEBS J., 2015

  • Voziyanova E., Malchin N., Anderson R.P., Yagil E., Kolot M., Voziyanov Y.
    Efficient Flp-Int HK022 dual RMCE in mammalian cells.
    Nucleic Acids Res., 2013

  • Anderson R.P., Voziyanova E., Voziyanov Y.
    Flp and Cre expressed from Flp-2A-Cre and Flp-IRES-Cre transcription units mediate the highest level of dual recombinase-mediated cassette exchange.
    Nucleic Acids Res., 2012

  • Shultz J.L., Voziyanova E., Konieczka J.H., Voziyanov Y.
    A genome-wide analysis of FRT-like sequences in the human genome.
    PLoS One, 2011

  • Malchin N., Molotsky T., Borovok I., Voziyanov Y., Kotlyar A.B., Yagil E., Kolot M.
    High efficiency of a sequential recombinase-mediated cassette exchange reaction in Escherichia coli.
    J. Mol. Microbiol. Biotechnol., 2010