Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
The human EF1a promoter does not provide expression of the transgene in mice. / Battulin, Nariman; Korablev, Alexey; Ryzhkova, Anastasia и др.
в: Transgenic Research, Том 31, № 4-5, 10.2022, стр. 525-535.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
}
TY - JOUR
T1 - The human EF1a promoter does not provide expression of the transgene in mice
AU - Battulin, Nariman
AU - Korablev, Alexey
AU - Ryzhkova, Anastasia
AU - Smirnov, Alexander
AU - Kabirova, Evelyn
AU - Khabarova, Anna
AU - Lagunov, Timofey
AU - Serova, Irina
AU - Serov, Oleg
N1 - Funding Information: This work was supported by Russian Foundation for Basic Research, grant number 20–04-60094. N.B. supported from the Ministry of Education and Science of Russian Federation, grant #2019–0546 (FSUS-2020–0040). Data analysis performed on computational nodes of the Institute of Cytology and Genetics (state project FWNR-2022–0019). Publisher Copyright: © 2022, The Author(s), under exclusive licence to Springer Nature Switzerland AG.
PY - 2022/10
Y1 - 2022/10
N2 - In this work, we set out to create mice susceptible to the SARS-CoV-2 coronavirus. To ensure the ubiquitous expression of the human ACE2 gene we used the human EF1a promoter. Using pronuclear microinjection of the transgene construct, we obtained six founders with the insertion of the EF1a-hACE2 transgene, from which four independent mouse lines were established. Unfortunately, only one line had low levels of hACE2 expression in some organs. In addition, we did not detect the hACE2 protein in primary lung fibroblasts from any of the transgenic lines. Bisulfite sequencing analysis revealed that the EF1a promoter was hypermethylated in the genomes of transgenic animals. Extensive analysis of published works about transgenic animals indicated that EF1a transgenic constructs are frequently inactive. Thus, our case cautions against using the EF1a promoter to generate transgenic animals, as it is prone to epigenetic silencing.
AB - In this work, we set out to create mice susceptible to the SARS-CoV-2 coronavirus. To ensure the ubiquitous expression of the human ACE2 gene we used the human EF1a promoter. Using pronuclear microinjection of the transgene construct, we obtained six founders with the insertion of the EF1a-hACE2 transgene, from which four independent mouse lines were established. Unfortunately, only one line had low levels of hACE2 expression in some organs. In addition, we did not detect the hACE2 protein in primary lung fibroblasts from any of the transgenic lines. Bisulfite sequencing analysis revealed that the EF1a promoter was hypermethylated in the genomes of transgenic animals. Extensive analysis of published works about transgenic animals indicated that EF1a transgenic constructs are frequently inactive. Thus, our case cautions against using the EF1a promoter to generate transgenic animals, as it is prone to epigenetic silencing.
KW - DNA methylation
KW - EF1a promoter
KW - Expression
KW - Pronuclear microinjection
KW - Transgenesis
KW - SARS-CoV-2/genetics
KW - Humans
KW - Mice, Transgenic
KW - Angiotensin-Converting Enzyme 2
KW - Animals
KW - COVID-19/genetics
KW - Mice
KW - Transgenes
UR - http://www.scopus.com/inward/record.url?scp=85135852633&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/33d814d2-25c6-3165-a872-747beb6c0256/
U2 - 10.1007/s11248-022-00319-5
DO - 10.1007/s11248-022-00319-5
M3 - Article
C2 - 35960480
AN - SCOPUS:85135852633
VL - 31
SP - 525
EP - 535
JO - Transgenic Research
JF - Transgenic Research
SN - 0962-8819
IS - 4-5
ER -
ID: 36916168