Evidence of Trangency and Humanization in Mice Obtained at the SCBMT of FMBA of Russia by Sanger Sequencing Method

Author:

Karkischenko N. N.1,Petrova N. V.1,Slobodenyuk V. V.1,Koloskova E. M.2,Laryushina N. A.1,Vasil’eva I. A.1,Petrov D. V.1,Bolotskih L. А.1,Savina M. A.1

Affiliation:

1. Scientific Center of Biomedical Technologies of the Federal Medical and Biological Agency of Russia

2. All-Russian Research Institute of Physiology, Biochemistry and Animal Nutrition — Branch of the Federal Scientific Centre of Animal Husbandry — All-Russian Institute of Animal Husbandry named after Acad. L.K. Ernst

Abstract

Several humanized transgenic lines of biomodel mice containing an integrated variable human gene of the main histocompatibility complex (MHC) have been created at the Federal State Budgetary Scientific and Scientific Research Institute of the FMBA of Russia. These include HLA-A*02:01, HLA-B*07:02 and HLA-C*07:02. The lines were created by microinjection of a linear fragment of a genetically engineered structure (GES) into the male pronucleus of zygotes, followed by the transfer of potentially modified embryos into the reproductive tract to pseudo-pregnant female recipients. The created GES encodes a chimeric molecule of the MHC of class I on the cell surface, consisting of the α1-, α2-domains of human HLA, and the α3-domain of the mouse H-2K complex, stabilized by human β2-microglobulin connected by a glycine serine linker with the α1-domain of HLA [1–5]. The created biomodels can be successfully used to solve a wide range of research tasks, including studies of immune reactions, infectious, autoimmune and oncological diseases, as well as the development and testing of vaccines in the field of pharmacosafety and immunogenicity. This article presents theoretical information on the genetic polymorphism of the studied gene in the human genome, as well as experimental data on the transgenic lines of biomodels created by the authors and the results of comparing the allele-specific site in the obtained animal lines. The analysis was performed by Sanger sequencing on a cDNA matrix.

Publisher

Scientific Center for Biomedical Technologies of the Federal Medical-Biological Agency

Reference16 articles.

1. Karkischenko V.N., Petrova N.V., Savchenko E.S., Ogneva N.S., Koloskova E.M., Maksimenko S.V., Manuvera V.A., Bobrovsky P.A., Lazarev V.N. Sozdanie polnih gibritnih DNK konstrukcij s genom cheloveka HLA-A*02:01:01:01 [Chimeric Construct Engineering with Human Variant HLA-A*02:01:01:01]. Biomeditsina [Journal Biomed]. 2021;17(1):10–23. (In Russian).

2. Karkischenko N.N., Lazarev V.N., Manuvera V.A., Bobrovsky P.A., Petrova N.V., Koloskova E.M., Glotova E.S. Principles of Creation of a Genetic Engineering Construction for Obtaining Humanized Transgenic Mice with HLA-C*07:02:01:01, as a Promote of Innovative Transgenic and Knockout Biomodels. Biomeditsina [Journal Biomed]. 2024;20(1):8–20. (In Russian)]. DOI: 10.33647/2074-5982-20-1-8-20.

3. Karkischenko N.N., Ryabykh V.P., Karkischenko V.N., Koloskova E.M. Sozdanie gumanizirovannykh myshey dlya farmakotoksikologicheskikh issledovaniy (uspekhi, neudachi I perspektivy) [Creation of humanized mice for pharmacotoxicological research (successes, failures and prospects)]. Biomeditsina [ Journal Biomed]. 2014;1(3):4–22. (In Russian)

4. Petrova N.V., Skripkina M.M. Osobennosti organizacii soderzhaniya i vyvedeniya transgennyh linij myshej v NCBMT FMBA Rossii [Maintenance and Breeding of Transgenic Mouse Strains at the Scientific Center of Biomedical Technologies of the FMBA of Russia]. Biomeditsina [ Journal Biomed]. 2021;17(3E):70–75. DOI: 10.33647/2713-0428-17-3E-70-75.

5. Savchenko E.S., Ogneva N.S., Karkischenko N.N. Embriologicheskie aspekty sozdaniya novoj gumanizirovannoj transgennoj linii myshej s integrirovannym genom cheloveka HLA-A*02:01:01:01 [Embryological aspects of creation a new humanized transgenic mice with integrated human HLA-A*02:01:01:01 gene]. Biomeditsina [Journal Biomed]. 2022;18(4):10–23. (In Russian)

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