universal-efficient-electroporation-protocol-for-genetic-engineering

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

Electroporation is a common method for transfection with different kinds of molecules by electrical permeabilization of the plasma membrane. With the ...

Technische Universität Dresden

The role of microRNAs (miRNA) in estrogen receptor (ER) signaling in the uterus and in endometrial cancer is not well understood. We therefore analyzed miRNA expression in uterine samples from a standard 3-day uterotrophic assay using young female adult rats to identify E2-regulated miRNAs. Microarray analysis identified 47 E2 down-regulated miRNAs including miR-30a, and 25 E2up-regulated miRNAs including miR-672, miR-203, and miR-146b. The strongly E2-upregulated miR-203 was selected for further analysis. miR-203 was deleted in the rat endometrial adenocarcinoma cell line, RUCA-I, using CRISPR/CAS9. Five clones devoid of miR-203 expression were generated. Proliferation was reduced and G2-arrest was observed in all miR-203 deficient RUCA-I clones. Transfection with a miR-203-3p mimic partially rescues this effect. Comparison of mRNA expression in three miR-203 knockout clones to wild type RUCA-I cells reveals 566 miR-203-upregulated and 592 miR-203-downregulated genes. 43 of the genes that are upregulated by miR-203 knockout in vitro are downregulated in the uterus by E2. Of these Acer2, Zbtb20, Ptn, Rcbtb2, Mum1l1, Hmgn3, and Nfat5 possess one or more seed sequence matches in their 3'-UTR that are predicted to be targets of miR-203. These data demonstrate the importance of E2 regulated miRNAs in general, and miR-203 in particular, for E2 regulated gene expression and physiological processes including proliferation and cell migration, in the uterus as well as in the etiology of endometrial carcinomas.

Role of miR-203 in estrogen receptor-mediated signaling in the rat uterus and endometrial carcinoma.

Organoid cultures of human pancreatic ductal adenocarcinoma (PDAC) have become a promising tool for tumor subtyping and individualized chemosensitivity testing. PDACs have recently been grouped into different molecular subtypes with clinical impact based on cytokeratin-81 (KRT81) and hepatocyte nuclear factor 1A (HNF1A). However, a suitable antibody for HNF1A is currently unavailable. The present study is aimed at establishing subtyping in PDAC organoids using an alternative marker.

CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids.

Patterns of genetic alterations characterize different molecular subtypes of human gastric cancer. We aimed to establish mouse models of these subtypes.

Mouse Models of Human Gastric Cancer Subtypes With Stomach-Specific CreERT2-Mediated Pathway Alterations.

[This corrects the article DOI: 10.1155/2019/1024614.].

Alexander Hennig

Department of Visceral-,
Thoracic and Vascular Surgery,
University Hospital Carl Gustav Carus,
Department of Visceral-, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus

Technische Universität Dresden

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

Alexander Hennig

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Visceral-,Thoracic and Vascular Surgery,University Hospital Carl Gustav Carus,Department of Visceral-, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus

Technische Universität Dresden

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

Department of Visceral-,
Thoracic and Vascular Surgery,
University Hospital Carl Gustav Carus,
Department of Visceral-, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus