Physiological and embryological aspects of generation transgenic mice with integrated human NAT1 and NAT2 genes

The method to induce superovulation in mice-donors, including the synchronization of sexual cycles of the female donor by pre-infusion to males through the partition, which allows you to get to determine time up to 17 zygotes per donor with highly visible pronucleus, suitable for introduction in them of genetically engineered structures was improved. Linear fragments of plasmid gene-engineering constructions comprising the nucleotide sequence of the human NAT1 and NAT2 genes under the promoter of the mouse albumin ( hNAT1 и hNAT2 ), microinjection into the male pronucleus of zygotes collected from the female F1 hybrid mice (CBA/lac * C57BL/6). In the preparation of the female recipients were also used taking prereplanting female recipients to vasectomy penis males through the partition, which contributed to the stimulation of the process of folliculogenesis and synchronization of sexual cycles in females-recipients. It is possible to obtain a greater number of female pseudoharmonic-recipients with copulation wads to determine time, which is one of the criteria of usefulness of the animal recipient and the possibility of successful transplantation of the embryos to it, microinjection genetically engineered constructs. Based on these techniques, developed variant of a modified technology for producing transgenic mice with the genes NAT1 and human NAT2 .

transgenic mouse NAT1 and NAT2, transplantation DNA constructs, folliculogenesis, modification of the technology for generation transgenic mice with human genes

1. Аллен Н. и др. Получение трансгенных мышей // Биология развития млекопитающих. Методы / под ред. М. Манк. - М.: Мир. 1990. С. 278-298.

2. Каркищенко Н.Н., Петрова Н.В., Слободенюк В.В. Высокоспецифичные видовые праймеры к генам NAT1 и NAT2 для сравнительных исследований у человека и лабораторных животных // Биомедицина. 2014. № 2. С. 4-17.

3. Каркищенко Н.Н., Рябых В.П., Каркищенко В.Н., Колоскова Е.М. Создание гуманизированных мышей для фармакотоксикологических исследований (успехи, неудачи и перспективы) // Биомедицина. 2014. № 3. С. 4-24.

4. Мерфи Д., Хенсон Дж. Получение трансгенных мышей путём микроинъекции клонированной ДНК в оплодотворённые яйцеклетки // Новое в клонировании ДНК. Методы / под ред. Д. Гловера. - М.: Мир. 1989. 386 с.

5. Boukouvala S., Price N., Sim E. Identification and functional characterization of novel polymorphisms associated with the genes for arylamine N-acetyltransferases in mice // Pharmacogenetics. 2002. No. 12. Pp. 385-394.

6. Butcher N.J., Arulpragasam A., Goh H.L., Davey T., Minchin R.F. Human arylamine N-acetyltransferase-1 interacts with EHZF, a multi-functional transcription co-factor // In: Third International Workshop on Arylamine N-acetyltransferases. -Vancouver, Canada. 2004.

7. De Leon J.H., Vatsis K.P., Weber W.W. Characterisation of naturally occuring and recombinant human N-actyltransferase variants encoded by NAT1* // Mol. pharmacol. 2000. No. 58. Pp. 288-299.

8. Estrada-Rodgers L., Levy G.N., Weber W.W. Substrate selectivity of mouse N-acetyltransferase 1, 2, and 3 expressed in COS-1 cells // Drug metab. dispos. 1998. No. 26. Pp. 502-505.

9. Fakis G., Boukouvala S., Buckle V., Payton M., Denning C., Sim E. Chromosomal localisation and mapping of the genes for murine arylamine N-acetyltransferases (NATs), enzymes involved in the metabolism of carcinogens: identification of a novel upstream non-coding exon for murine Nat2 // Cytogenet. cell genet. 2000. No. 90. Pp. 134-138.

10. Fretland A., Doll M.A., Gray K., Fen Y., Hein D. Cloning, sequencing and recombinant expression of NAT1, NAT2 and NAT3 derived from the C3H/HeJ (rapid) and A/HeJ (slow) acetylator inbred mouse: functional characterisation of the activation and deactivation of aromatic amine carcinogens // Toxicol. appl. pharmacol. 1997. No. 142. Pp. 360-366.

11. Glowinski I.B., Weber W.W. Biochemical characterisation of genetically variant aromatic amine N-acetyltransferases in A/J and C57BL/6J mice // J. biol. chem. 1982. No. 257. Pp. 1431-1437.

12. Hein D.W., Trinidad A., Yerokun T., Ferguson R.J., Kirlin W.G., Weber W.W. Genetic control of acetyl coenzyme A-dependent arylamine N-acetyltransferase, hydrazine N-acetyltransferase, and N-hydroxy-arylamine O-acetyltransferase enzymes in C57BL/6J, A/J, AC57F1, and the rapid and slow acetylator A.B6 and B6.A congenic inbred mouse // Drug metab. dispos. 1988. No. 16. Pp. 341-347.

13. Karolyi J., Erickson R.P., Liu S. Genetics of susceptibility to 6-aminonicotinamide-induced cleft palate in the mouse: studies in congenic and recombinant inbred strains // Teratology. 1988. No. 37. Pp. 283-287.

14. Karolyi J., Erickson R.P., Liu S., Killewald L. Major effects on teratogen-induced facial clefting in mice determined by a single genetic region // Genetics. 1990. No. 126. Pp. 201-205.

15. Kelly S.L., Sim E. Arylamine N-acetyltransferase in Balb/c mice: identification of a novel mouse isoenzyme by cloning and expression in vitro // Biochem. J. 1994. No. 302. Pp. 347-353.

16. Martell K.J., Levy G.N., Weber W.W. Cloned mouse N-acetyltransferases: enzymatic properties of expressed Nat-1 and Nat-2 gene products // Mol. pharmacol. 1992. No. 42. Pp. 265-272.

17. Sakurai T., Kimura M., Sato M. Temporary developmental arrest after storage of fertilized mouse oocytes at 4C: effects on embryonic development, maternal mRNA processing and cell cycle // Molecular human reproduction. 2005. Vol. 11. No. 5. Pp. 325-333.

18. Sakurai T., Watanabe S., Kamiyoshi A., Sato M., Shindo T. A single blastocyst assay optimized for detecting CRISPR/Cas9 system-induced indel mutations in mice // BMC Biotechnology. 2014. No. 14. Pp. 69-79.