INVESTIGATIONS ON EPITHELIAL BIOLOGY USING ORGANOID DIFFERENTIATION AND CO-CULTURES

10.33540/182 ◽  
2021 ◽  
Author(s):  
◽  
Jens Puschhof
Keyword(s):  
Epithelial Biology

scholarly journals Functional Genomic Annotation of Genetic Risk Loci Highlights Inflammation and Epithelial Biology Networks in CKD

2014 ◽  
Vol 26 (3) ◽  
pp. 692-714 ◽  
Author(s):  
Nora Ledo ◽  
Yi-An Ko ◽  
Ae-Seo Deok Park ◽  
Hyun-Mi Kang ◽  
Sang-Youb Han ◽  
...  
Keyword(s):  
Genetic Risk ◽  
Functional Genomic ◽  
Genomic Annotation ◽  
Epithelial Biology

scholarly journals Knock-in tagging in zebrafish facilitated by insertion into non-coding regions

2021 ◽  
Author(s):  
Daniel S Levic ◽  
Naoya Yamaguchi ◽  
Siyao Wang ◽  
Holger Knaut ◽  
Michel Bagnat
Keyword(s):  
Protein Function ◽  
Cell Biology ◽  
Quantitative Imaging ◽  
Expression Patterns ◽  
Imaging Studies ◽  
Coding Regions ◽  
Fluorescent Markers ◽  
Endogenous Expression ◽  
Epithelial Biology

Zebrafish provide an excellent model for in vivo cell biology studies due to their amenability to live imaging. Protein visualization in zebrafish has traditionally relied on overexpression of fluorescently tagged proteins from heterologous promoters, making it difficult to recapitulate endogenous expression patterns and protein function. One way to circumvent this problem is to tag the proteins by modifying their endogenous genomic loci. Such an approach is not widely available to zebrafish researchers due to inefficient homologous recombination and the error-prone nature of targeted integration in zebrafish. Here, we report a simple approach for tagging proteins in zebrafish on their N- or C termini with fluorescent markers by inserting PCR-generated donor amplicons into non-coding regions of the corresponding genes. Using this approach, we generated endogenously tagged alleles for several genes critical for epithelial biology and organ development including the tight junction components ZO-1 and Cldn15la, the trafficking effector Rab11a, and the ECM receptor β1 integrin. Our approach facilitates the generation of knock-in lines in zebrafish, opening the way for accurate quantitative imaging studies.

scholarly journals p63 – The Master Key to Epithelial Biology

2018 ◽  
Vol 5 (8) ◽  
Author(s):  
Anitha Dayakar ◽  
Pushparaja Shetty ◽  
Mundoor Manjunath Dayakar
Keyword(s):  
Epithelial Biology

scholarly journals A semi-automated organoid screening method demonstrates epigenetic control of intestinal epithelial differentiation

2020 ◽  
Author(s):  
Jenny Ostrop ◽  
Rosalie Zwiggelaar ◽  
Marianne Terndrup Pedersen ◽  
François Gerbe ◽  
Korbinian Bösl ◽  
...  
Keyword(s):  
Therapeutic Potential ◽  
Screening Method ◽  
Adult Stem Cell ◽  
Epithelial Differentiation ◽  
Type I ◽  
Intestinal Epithelial ◽  
Specific Chemical ◽  
Organoid Cultures ◽  
Resource Data ◽  
Epithelial Biology

AbstractIntestinal organoids are an excellent model to study epithelial biology. Yet, the selection of analytical tools to accurately quantify heterogeneous organoid cultures remains limited. Here, we developed a semi-automated organoid screening method, which we applied to a library of highly specific chemical probes to identify epigenetic regulators of intestinal epithelial biology. The role of epigenetic modifiers in adult stem cell systems, such as the intestinal epithelium, is still undefined. Based on this resource data, we identified several targets that affected epithelial cell differentiation, including HDACs, EP300/CREBBP, LSD1, and type I PRMTs, which were verified by complementary methods. For example, we show that inhibiting type I PRMTs, which leads enhanced epithelial differentiation, blocks the growth of adenoma but not normal organoid cultures. Thus, epigenetic probes are powerful tools to study intestinal epithelial biology and may have therapeutic potential.

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