The human EF1a promoter does not provide expression of the transgene in mice.

Content Provider	Europe PMC
Author	Battulin, Nariman Korablev, Alexey Ryzhkova, Anastasia Smirnov, Alexander Kabirova, Evelyn Khabarova, Anna Lagunov, Timofey Serova, Irina Serov, Oleg
Abstract	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.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11248-022-00319-5.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC9372930&blobtype=pdf
ISSN	09628819
Journal	Transgenic Research [Transgenic Res]
Volume Number	31
DOI	10.1007/s11248-022-00319-5
PubMed Central reference number	PMC9372930
Issue Number	4-5
PubMed reference number	35960480
e-ISSN	15739368
Language	English
Publisher	Springer International Publishing
Publisher Date	2022-08-12
Publisher Place	Gewerbestrasse 11, Cham, Ch 6330, Switzerland
Access Restriction	Open
Rights License	This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. © The Author(s), under exclusive licence to Springer Nature Switzerland AG 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
Subject Keyword	EF1a promoter Transgenesis Pronuclear microinjection DNA methylation Expression
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science