Development of the lentiviral vector with GC-rich segment of the promoter region of fusion oncogene RUNX1-RUNX1T1

Authors

  • Hanna A. Saurytskaya Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus
  • Ilya M. Ilyushonak Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus
  • Vasily V. Grinev Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

Keywords:

fusion oncogene RUNX1-RUNX1T1, molecular cloning, GC-rich region, polymerase chain reaction
Supporting Agencies
Authors are sincerely grateful to Alexandr Migas (Department of Research, Belarusian Research Center for Pediatric Oncology, Hematology and Immunology, Minsk, Belarus) for assistance in Sanger sequencing. Authors H. A. Saurytskaya, I. M. Ilyushonak contribution is equal.

Abstract

As is known, a PCR-cloning of GC-rich regions and development the vectors with such sequences can be non trivial. In this work we performed an optimization of PCR-amplification protocol for segment of internal promoter region of fusion oncogene RUNX1-RUNX1T1 and developed of multipurpose lentiviral vector with insertion of this sequence. We generalize our experience of dealing with GC-rich sequences in some methodological recommendations.

Author Biographies

  • Hanna A. Saurytskaya, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

    student at the faculty of biology

  • Ilya M. Ilyushonak, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

    assistant at the department of genetics, faculty of biology

  • Vasily V. Grinev, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

    PhD (biology); associate professor at the department of genetics, faculty of biology

References

  1. Döhner H., Weisdorf D. J., Bloomfield C. D. Acute Myeloid Leukemia. N. Engl. J. Med. 2015. Vol. 373, No. 12. P. 1136 –1152.
  2. Müller A. M. S., Duque J., Shizuru J. A., et al. Complementing mutations in core binding factor leukemias: from mouse models to clinical applications. Oncogene. 2008. Vol. 27, No. 44. P. 5759 –5773.
  3. Basecke J., Cepek L., Mannhalter C., et al. Transcription of AML1/ETO in bone marrow and cord blood of individuals without acute myelogenous leukemia. Blood. 2002. Vol. 100, No. 6. P. 2267–2268.
  4. Mori H., Colman S. M., Xiao Z., et al. Chromosome translocations and covert leukemic clones are generated during normal fetal development Proc. Natl. Acad. Sci. USA. 2002. Vol. 99, No. 12. P. 8242–8247.
  5. Markova E. N., Kantidze O. L., Razin S. V. Transcription of the AML1/ETO chimera is guided by the P2 promoter of the AML1 gene in the Kasumi-1 cell line. Gene. 2012. Vol. 510, No. 2. P. 142–146.
  6. Migas A. A., Mishkova O. A., Ramanouskaya T. V., et al. RUNX1T1/MTG8/ETO gene expression status in human t (8; 21) (q22; q22)-positive acute myeloid leukemia cells. Leuk. Res. 2014. Vol. 38, No. 9. P. 1102–1110.
  7. Sambrook J., Russel W. Preparation and analysis of eukaryotic genomic DNA. Molecular cloning. A laboratory manual. New York : Cold Spring Harbor, 2001. Vol. 1. P. 524 –580.
  8. Gupta R., Wikramasinghe P., Bhattacharyya A., et al. Annotation of gene promoters by integrative data-mining of ChIP-seq Pol-II enrichment data. BMC Bioinformatics. 2010. 11 (Suppl. 1): S65. DOI: 10.1186/1471-2105-11-S1-S65.
  9. Levanon D., Glusman G., Bangsow T., et al. Architecture and anatomy of the genomic locus encoding the human leukemia-associated transcription factor RUNX1/AML1. Gene. 2001. Vol. 262. P. 23–33.
  10. Ghozi M. C., Bernstein Y., Negreanu V., et al. Expression of the human acute myeloid leukemia gene AML1 is regulated by two promoter regions. Proc. Natl. Acad. Sci. USA. 1996. Vol. 93, No. 5. P. 1935–1940.
  11. Grinev V. V., Radzisheuskaya A. A., Heidenreich O. Effective transgene transfer to human leukemic cells with the second generation lentiviral vector. Vestnik BGU. Ser. 2, Khim. Biol. Geogr. 2008. No. 3. P. 82– 86 (in Russ.).

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Published

2018-05-02

Issue

No. 3 (2017): Journal of the Belarusian State University. Biology

Section

Genetics and Molecular Biology

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How to Cite

Saurytskaya, H. A., Ilyushonak, I. M., & Grinev, V. V. (2018). Development of the lentiviral vector with GC-rich segment of the promoter region of fusion oncogene RUNX1-RUNX1T1. Experimental Biology and Biotechnology, 3, 38-44. https://journals.bsu.by/index.php/biology/article/view/2459