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用于细胞特异性分析的EKLF(Klf1)/ CRE基因敲入小鼠的构建、表征及应用

Generation, characterization, and use of EKLF(Klf1)/CRE knock-in mice for cell-restricted analyses.

作者信息

Xue Li, Mukherjee Kaustav, Kelley Kevin A, Bieker James J

机构信息

Department of Cell, Developmental, and Regenerative Biology, Mount Sinai School of Medicine, New York, NY, United States.

Black Family Stem Cell Institute, Mount Sinai School of Medicine, New York, NY, United States.

出版信息

Front Hematol. 2023;2. doi: 10.3389/frhem.2023.1292589. Epub 2024 Jan 19.

DOI:10.3389/frhem.2023.1292589
PMID:39280931
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11393758/
Abstract

INTRODUCTION

EKLF/Klf1 is a tissue-restricted transcription factor that plays a critical role in all aspects of erythropoiesis. Of particular note is its tissue-restricted pattern of expression, a property that could prove useful for expression control of a linked marker or enzymatic gene.

METHODS AND RESULTS

With this in mind, we fused the CRE recombinase to the genomic EKLF coding region and established mouse lines. We find by FACS analyses that CRE expression driven by the EKLF transcription unit recapitulates erythroid-restricted expression with high penetrance in developing embryos. We then used this line to test its properties in the adult, where we found EKLF/CRE is an active and is a robust mimic of normal EKLF expression in the adult bone marrow. EKLF/CRE is also expressed in erythroblastic island macrophage in the fetal liver, and we demonstrate for the first time that, as seen during embryonic development, EKLF is also expressed in adult BM-derived erythroblastic island macrophage. Our data also support lineage studies showing EKLF expression at early stages of hematopoiesis.

DISCUSSION

The EKLF/CRE mouse lines are novel reagents whose availability will be of great utility for future experiments by investigators in the red cell field.

摘要

引言

EKLF/Klf1是一种组织特异性转录因子,在红细胞生成的各个方面都发挥着关键作用。特别值得注意的是其组织特异性表达模式,这一特性可能对连接标记物或酶基因的表达控制有用。

方法与结果

考虑到这一点,我们将CRE重组酶与基因组EKLF编码区融合并建立了小鼠品系。通过流式细胞术分析,我们发现由EKLF转录单元驱动的CRE表达在发育中的胚胎中以高穿透率重现了红细胞特异性表达。然后我们使用该品系在成年小鼠中测试其特性,发现EKLF/CRE是活跃的,并且是成年骨髓中正常EKLF表达的有力模拟物。EKLF/CRE也在胎儿肝脏的成红细胞岛巨噬细胞中表达,并且我们首次证明,正如在胚胎发育过程中所见,EKLF也在成年骨髓来源的成红细胞岛巨噬细胞中表达。我们的数据还支持谱系研究,显示EKLF在造血早期阶段表达。

讨论

EKLF/CRE小鼠品系是新型试剂,其可用性将对红细胞领域的研究人员未来的实验非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b54b/11393758/2699b4e1c32b/nihms-1969873-f0006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b54b/11393758/305536833db0/nihms-1969873-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b54b/11393758/d5bd5d253443/nihms-1969873-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b54b/11393758/1c013070f10e/nihms-1969873-f0003.jpg
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