Effects of Tamoxifen Inducible MerCreMer on Gene Expression in Cardiac Myocytes in Mice

Authors

Leila Rouhi
Siyang Fan
Sirisha M Cheedipudi
Melis Olcum
Hyun-Hwan Jeong
Zhongming Zhao
Priyatansh Gurha
Ali J Marian

Publication Date

1-1-2022

Journal

The Journal of Cardiovascular Aging

Abstract

The Cre-LoxP technology, including the tamoxifen (TAM) inducible MerCreMer (MCM), is increasingly used to delineate gene function, understand the disease mechanisms, and test therapeutic interventions. We set to determine the effects of TAM-MCM on cardiac myocyte transcriptome. Expression of the MCM was induced specifically in cardiac myocytes upon injection of TAM to myosin heavy chain 6-MCM (Myh6-Mcm) mice for 5 consecutive days. Cardiac function, myocardial histology, and gene expression (RNA-sequencing) were analyzed 2 weeks after TAM injection. A total of 346 protein coding genes (168 up- and 178 down-regulated) were differentially expressed. Transcript levels of 85 genes, analyzed by a reverse transcription-polymerase chain reaction in independent samples, correlated with changes in the RNA-sequencing data. The differentially expressed genes were modestly enriched for genes involved in the interferon response and the tumor protein 53 (TP53) pathways. The changes in gene expression were relatively small and mostly transient and had no discernible effects on cardiac function, myocardial fibrosis, and apoptosis or induction of double-stranded DNA breaks. Thus, TAM-inducible activation of MCM alters cardiac myocytes gene expression, provoking modest and transient interferon and DNA damage responses without exerting other discernible phenotypic effects. Thus, the effects of TAM-MCM on gene expression should be considered in discerning the bona fide changes that result from the targeting of the gene of interest.

Keywords

Transcriptome, gene expression, tamoxifen, Cre recombinase, MerCreMer, interferon, inflammation, TP53

DOI

10.20517/jca.2021.30

PMID

35079750

PMCID

PMC8785140

PubMedCentral® Posted Date

1-24-2022

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

yes