Generation of H11-albumin-rtTA Transgenic Mice: A Tool for Inducible Gene Expression in the Liver

Abstract

The modification of the mouse genome by site-specific gene insertion of transgenes and other genetic elements allows the study of gene function in different developmental stages and in the pathogenesis of diseases. Here, we generated a “genomic safe harbor” Hipp11 ( H11 ) locus-specific knock-in transgenic mouse line in which the albumin promoter is used to drive the expression of the reverse tetracycline transactivator (rtTA) in the liver. The newly generated H11-albumin-rtTA transgenic mice were bred with tetracycline-operator-Histone-2B-green fluorescent protein (TetO-H2BGFP ) mice to assess inducibility and tissue-specificity. Expression of the H2BGFP fusion protein was observed exclusively upon doxycycline (Dox) induction in the liver of H11-albumin-rtTA/TetO-H2BGFP double transgenic mice. To further analyze the ability of the Dox-inducible H11-albumin-rtTA mice to implement conditional DNA recombination, H11-albumin-rtTA transgenic mice were crossed with TetO-Cre and Ai14 mice to generate H11-albumin-rtTA/TetO-Cre/Ai14 triple transgenic mice. We successfully confirmed that the Cre-mediated recombination efficiency was as strong in Dox-induced H11-albumin-rtTA /TetO-Cre/Ai14 mice as in the control albumin-Cre/A14 mice. Finally, to characterize the expression-inducing effects of Dox in H11-albumin-rtTA/TetO-H2BGFP mice in detail, we examined GFP expression in embryos at different developmental stages and found that newly conceived H11-albumin-rtTA/TetO-H2BGFP embryos of Dox-treated pregnant female mice were expressing reporter GFP by E16.5. Our study demonstrates that these new H11-albumin-rtTA transgenic mice are a powerful and efficient tool for the temporally and spatially conditional manipulation of gene expression in the liver, and illustrates how genetic crosses with these new mice enable the generation of complex multi-locus transgenic animals for mechanistic studies.

Proteomic analysis of coarse and fine skin tissues of Liaoning cashmere goat

“…In Malhotra's experiment, it was found that siRNA gene therapy with an albumin carrier, inhibit the expression of the genes involved to treat cancer [21]. In Li's experiment, the new H11-albumin-rTTA transgenic mice were found to be a powerful and effective tool for manipulating gene expression in the liver in both spatial and temporal conditions [22]. Lorraine's study evaluated the potential value of albuminuria as a "biomarker" for acute kidney injury (AKI) and examined whether AKI induces expression of normally silenced albumin genes in the kidney.…”

Comparative study on the expression characteristics of transgenes inserted into the <italic>Gt(ROSA)26Sor</italic> and H11 <italic>loci</italic> in mice

“…It is situated between the Eif4enif1 gene and the Drg1 gene on chromosome 11, with no endogenous genes identified within this region. Several transgenic models inserted at this locus have confirmed its safety as a harbor [12][13][14].…”

Spermatogenic cell-specific type 1 hexokinase (HK1S) is essential for capacitation-associated increase in tyrosine phosphorylation and male fertility in mice

“…Standard Hematoxylin and Eosin (H&E) staining was performed on paraffin sections as described previously [ 47 ]. Testes and cauda epididymides were freshly fixed in Davidson’s fixative solution (Formaldehyde: Ethanol: Glacial acetic acid: H 2 O = 6: 3: 1: 10) overnight at 4°C.…”