Characterization of the adiponectin promoter + Cre recombinase insertion in the Tg(Adipoq-cre)1Evdr mouse by targeted locus amplification and droplet digital PCR
| Autor: | Adrian M. Wong, Tushar P. Patel, Elizabeth K. Altman, Nicol Tugarinov, Giampaolo Trivellin, Jack A. Yanovski |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Adipocyte, Vol 10, Iss 1, Pp 21-27 (2021) |
| Druh dokumentu: | article |
| ISSN: | 2162-3945 2162-397X 21623945 |
| DOI: | 10.1080/21623945.2020.1861728 |
| Popis: | The Tg(Adipoq-cre)1Evdr mouse has become an important tool in adipose tissue biology. However, the exact genomic transgene integration site has not been established. Using Targeted Locus Amplification (TLA) we found the transgene had integrated on mouse chromosome 9 between exons 6 and 7 of Tbx18. We detected transgene-transgene fusion; therefore, we used droplet digital polymerase chain reaction to identify Cre copy number. In two separate experiments, we digested with BAMHI and with HindIII to separate potentially conjoined Cre sequences. We found one copy of intact Cre present in each experiment, indicating transgene-transgene fusion in other parts of the BAC that would not contribute to tissue-specific Cre expression. Cre copy number for Tg(Adipoq-cre)1Evdr mice can be potentially used to identify homozygous mice. |
| Databáze: | Directory of Open Access Journals |
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