Identification of hematopoietic-specific regulatory elements from the CD45 gene and use for lentiviral tracking of transplanted cells.
Identification of hematopoietic-specific regulatory elements from the CD45 gene and use for lentiviral tracking of transplanted cells.
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DOI：
10.1016/j.exphem.2014.05.005
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发表时间：
2014-09
影响因子：
2.6
通讯作者：
Levasseur, Dana N.
中科院分区：
文献类型：
--
作者：
Duong, Khanh L.;Das, Satyabrata;Yu, Shuyang;Barr, Jennifer Y.;Jena, Snehalata;Kim, Eunmi;Zavazava, Nicolas;Colgan, John D.;Xue, Hai-Hui;Levasseur, Dana N.
造血报告基因的开发对于确定源自基于细胞的疗法的谱系的命运至关重要。标记系统将能够实现更安全的基于胚胎干（ES）和诱导多能干（iPS）细胞的血液谱系衍生，并促进基于直接成纤维细胞转化的有效细胞重编程策略的开发。在这里，我们报告的蛋白酪氨酸磷酸酶CD 45是一个理想的候选基因的基础上，造血报告。通过分析转录因子染色质占有率（ChIP-seq）和启动子核酸酶敏感性（DNase-seq）发现了CD 45调控元件，以鉴定表达所需的最低限度足够序列。在将CD 45调控元件克隆到减毒慢病毒骨架中后，我们发现两个转录起始区对于高水平表达是必需的。在原代B细胞分化试验和移植模型中表达含有这些区域并与GFP连接的CD 45启动子导致小鼠和人血细胞谱系中淋巴样、髓样和有核红细胞中高水平的GFP。此外，在二次移植后5个月仍保持高GFP水平，表明报告基因的持久性。在成纤维细胞或ES细胞转导后没有观察到⑶ 45驱动的GFP表达。GFP报告基因仅在ES细胞分化为造血祖细胞和谱系后才能看到，这表明这种造血报告基因系统可用于验证潜在的自体血细胞疗法。
The development of a hematopoietic reporter is crucial for determining the fate of lineages derived from cell-based therapies. A marking system will enable safer embryonic stem (ES) and induced pluripotent stem (iPS) cell-based derivation of blood lineages, and facilitate the development of efficient cellular reprogramming strategies based on direct fibroblast conversion. Here we report that the protein tyrosine phosphatase CD45 is an ideal candidate gene on which to base a hematopoietic reporter. CD45 regulatory elements were discovered by analyzing transcription factor chromatin occupancy (ChIP-seq) and promoter nuclease sensitivity (DNase-seq) to identify minimally sufficient sequences required for expression. After cloning the CD45 regulatory elements into an attenuated lentiviral backbone, we found that two transcriptional initiation regions were essential for high-level expression. Expressing CD45 promoters containing these regions and tethered to GFP in a primary B cell differentiation assay and a transplantation model resulted in high levels of GFP in lymphoid, myeloid and nucleated erythroid cells in mouse and human blood cell lineages. Moreover, high GFP levels remained five months following secondary transplantation, indicating persistence of the reporter. No CD45 driven GFP expression is observed following fibroblast or ES cell transduction. The GFP reporter is seen only after ES cells differentiate into hematopoietic cell progenitors and lineages, suggesting that this hematopoietic reporter system could be useful in validating potential autologous blood cell therapies.
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DOI：
10.1186/1745-7580-4-5
发表时间：
2008-04-29
期刊：
Immunome research
影响因子：
--
作者：
Lattin JE;Schroder K;Su AI;Walker JR;Zhang J;Wiltshire T;Saijo K;Glass CK;Hume DA;Kellie S;Sweet MJ
通讯作者：
Sweet MJ
DOI：
10.1084/jem.20110447
发表时间：
2011-11-21
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kataoka K;Sato T;Yoshimi A;Goyama S;Tsuruta T;Kobayashi H;Shimabe M;Arai S;Nakagawa M;Imai Y;Kumano K;Kumagai K;Kubota N;Kadowaki T;Kurokawa M
通讯作者：
Kurokawa M
影响因子：
20.3
作者：
Chan, Kun-Ming;Bonde, Sabrina;Zavazava, Nicholas
通讯作者：
Zavazava, Nicholas
影响因子：
23.9
作者：
Rossi, Lara;Lin, Kuanyin K.;Boles, Nathan C.;Yang, Liubin;King, Katherine Y.;Jeong, Mira;Mayle, Allison;Goodell, Margaret A.
通讯作者：
Goodell, Margaret A.
影响因子：
64.8
作者：
Robinton, Daisy A.;Daley, George Q.
通讯作者：
Daley, George Q.