构建flattp - t2a - h2b - venus x C-peptide-mCherry双报告细胞系，监测WNT/Planar细胞极性通路活性

IF 0.7 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Stem cell research Pub Date : 2025-09-19 DOI:10.1016/j.scr.2025.103838

Tobias Greisle , Ines Kunze , Xianming Wang , Andrzej R. Malinowski , Anika Böttcher , Heiko Lickert , Ingo Burtscher

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引用次数: 0

摘要

从人诱导多能干细胞（hiPSCs）中获得功能性β细胞，在糖尿病研究中具有细胞替代治疗、疾病建模和药物测试的潜力。Wnt/平面细胞极性（PCP）信号在小鼠内分泌细胞发育和β细胞成熟中起着至关重要的作用，可以通过组织特异性效应基因Flattop的表达进行追踪。在这里，我们报道了通过CRISPR/Cas9基因编辑产生的人类荧光FLTP/CFAP126 （Flattop-T2A-H2B-Venus）和FLTP- insulin （Flattop-T2A-H2B-Venus x C-peptide-mCherry）双报告基因。这些hiPSC报告系可以监测内分泌细胞形成过程中的WNT/PCP信号，并研究其在人类模型系统中β细胞中的作用。

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Generation of a Flattop-T2A-H2B-Venus x C-peptide-mCherry double reporter human iPSC line to monitor WNT/Planar cell polarity pathway activity

Deriving functional β-cells from human induced pluripotent stem cells (hiPSCs) holds potential for cell replacement therapy, disease modeling, and drug testing in diabetes research. Wnt/Planar cell polarity (PCP) signaling is crucial for endocrine cell development and β-cell maturation in murine models and can be tracked by the expression of the tissue-specific effector gene Flattop. Here, we report the generation of a human fluorescent FLTP/CFAP126 (Flattop-T2A-H2B-Venus) and FLTP-Insulin (Flattop-T2A-H2B-Venus x C-peptide-mCherry) double reporter by CRISPR/Cas9 gene editing. These hiPSC reporter lines allow monitoring of WNT/PCP signaling during endocrine cell formation and studying its role in β-cells in a human model system.

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来源期刊

Stem cell research 生物-生物工程与应用微生物

CiteScore

2.20

自引率

8.30%

发文量

338

审稿时长

55 days

期刊介绍： Stem Cell Research is dedicated to publishing high-quality manuscripts focusing on the biology and applications of stem cell research. Submissions to Stem Cell Research, may cover all aspects of stem cells, including embryonic stem cells, tissue-specific stem cells, cancer stem cells, developmental studies, stem cell genomes, and translational research. Stem Cell Research publishes 6 issues a year.