Dr. Mikhail Kolot

ביוכימיה קלינית סגל אקדמי בכיר
Dr. Mikhail Kolot
Phone: 03-6406695
Fax: 03-6406834
Office: Sherman - Life Sciences, 611

Research Interests

HK022 bacteriophage site specific recombinase Integrase is the tool for human genome manipulation and gene therapy. Site specific recombinases (SSRs) catalase the site-specific recombination reaction between two specific short DNA sequences of 30-40 bp that serve as recombination sites that included integration, excision and inversion. These enzymes are widely used for the genome manipulations and gene therapy. E. coli HK022 bacteriophage site specific recombinase Integrase (HK-Int) belongs to the tyrosine family of SSRs and catalyzes the phage integration into E. coli chromosome and prophage excision. We have previously adopted the HK-Int for genome manipulation in plants, Cyanobacteria and human cells.

 

We have found that the HK-Int can utilize endogenous human attB sites that flank human deleterious mutations. The main goal of this research is to adopt the HK-Int for curing human hereditary diseases using the Recombinase Mediated Cassette Exchange reaction.  We expect that our research will lead to the development of novel efficient human genome manipulation technology that will be useful for gene therapy of hereditary diseases. 

 

We have developed a new cancer specific cells killing binary system activated by the HK- Int. We have validated the cancer cells killing activity of our new binary system in LLC1 Katushka lung cancer mice using a diphtheria toxin gene. We expect that our system will be used in the future for the establishment of a specific cancer cells killing system for lung cancer, as well as for other different types of cancer.

 

Recent Publications

Zaritsky, A., Ben-Dov, E., Borovsky, D., Boussiba, S., Einav, M., Gindin, G., Horowitz, R. Kolot, M., Melnikov, O. and Yagil, E. (2010) Transgenic organisms expressing genes from Bacillus thurigiensis to combat insect pests. Bioengineered Bugs 1:341-344.

 

Malchin, N., Molotsky, T., Borovok, I., Voziyanov, Y., Kotlyar A.B., Yagil, E., Kolot, M*. (2010) High Efficiency of a Sequential Recombinase-Mediated Cassette Exchange Reaction in E. coli. J. Molec. Microbiol. and Biotechnol. 19:117-122.

 

Malchin, N, Tuby, C.N., Yagil, E. and Kolot M*. (2011) Arm site independence of coliphage HK022 integrase activity in human cells.  Molec. Genet. Genomics 285: 403-413.

 

Keren-Kaplan T, Attali I, Motamedchaboki K, Davis BA, Tanner N, Reshef Y, Laudon E, Kolot M, Levin-Kravets O, Kleifeld O, Glickman M, Horazdovsky BF, Wolf DA, Prag G. (2011) Synthetic biology approach to reconstituting the ubiquitylation cascade in bacteria. EMBO 31(2):378-90.

 

Moshe, A., Pfannstiel, J., Yariv, B., Kolot, M., Sobol, I., Czosnek, H. and Gorovits, R. (2012) Stress responses to Tomato Yellow Leaf Curl Virus (TYLCV) infection of resistant and susceptible tomato plants are different. Metabolomics dx.doi.org/10.4172/2153-0769.S1-006.

 

Gorovits, R., Moshe, A., Kolot, M., Sobol, I., Czosnek, H. (2012) Progressive aggregation of Tomato yellow leaf curl virus coat protein in   systemically infected tomato plants, susceptible and resistant to the virus. Virus Research doi:10.1016/j.virusres.2012.09.017

 

Voziyanova, E. Malchin, N., Anderson, R.P., Yagil, E., Kolot, M*.,Voziyanov, Y*. (2013) Efficient Flp-Int HK022 dual RMCE in mammalian cells. Nucl. Acid Res. 1; 41(12):e125. doi: 10.1093/nar/gkt341

 

Kolot M, Malchin N, Elias A, Gritsenko N, Yagil E. (2015)  Site promiscuity of coliphage HK022 integrase as a tool for gene therapy. Gene Therapy. doi: 10.1038/gt.2015.9

 

Yagil E., Kolot M. (2015) Microbial site-specific recombinases as gene editing tools. J. Bacteriol. Mycol. Open Access 1(1): 00005. DOI: 10.15406/jbmoa.2015.01.00005

 

Rotem O., Nesper J., Borovok I., Gorovits K., Kolot M., Pasternak Z., Shin I., Glatter T., Pietrokovski S., Jenal U., Jurkevitch E. (2015) An extended cyclic di-GMP network in the predatory bacterium Bdellovibrio bacteriovorus. J. Bacteriol. pii: JB.00422-15

 

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