Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells.
Lentiviral vectors containing an enhancer-less ubiquitously acting chromatin opening element (UCOE) provide highly reproducible and stable transgene expression in hematopoietic cells.
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DOI：
10.1182/blood-2006-12-060814
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发表时间：
2007-09-01
期刊：
影响因子：
20.3
通讯作者：
Thrasher AJ
中科院分区：
文献类型：
--
作者：
Zhang F;Thornhill SI;Howe SJ;Ulaganathan M;Schambach A;Sinclair J;Kinnon C;Gaspar HB;Antoniou M;Thrasher AJ
泛在作用的染色质开放元件（UCOE）由无甲基化的CpG岛组成，所述CpG岛包含管家基因的双趋异转录启动子，所述管家基因已显示赋予对转录沉默的抗性并在组织培养系统中产生一致且稳定的转基因表达。为了开发用于造血细胞基因治疗的改进策略，我们评估了新型人HNRPA 2B 1-CBX 3 UCOE（A2 UCOE）在自失活（SIN）慢病毒载体背景下的潜力。与病毒启动子不同，无增强子的A2 UCOE产生了以高度可重复的水平表达报告基因转基因的细胞群体。每个载体基因组的表达效率也显着增加，在体内相比，载体纳入脾病灶形成病毒（SFFV）或巨细胞病毒（CMV）启动子，表明沉默的相对抗性。此外，A2 UCOE-IL 2 RG载体在体外完全恢复了IL 2 RG缺陷型人细胞内的IL-2信号传导途径，并在该疾病的小鼠模型中成功挽救了X连锁严重联合免疫缺陷（SCID-X1）表型。这些数据表明，A2 UCOE在慢病毒载体内显示出高度可靠的转录活性，在很大程度上克服了插入位点位置效应，并产生了治疗相关的基因表达水平。这些特性是在不存在经典增强子活性的情况下实现的，因此可以赋予高安全性特征。
Ubiquitously acting chromatin opening elements (UCOEs) consist of methylation-free CpG islands encompassing dual divergently transcribed promoters of housekeeping genes that have been shown to confer resistance to transcriptional silencing and to produce consistent and stable transgene expression in tissue culture systems. To develop improved strategies for hematopoietic cell gene therapy, we have assessed the potential of the novel human HNRPA2B1-CBX3 UCOE (A2UCOE) within the context of a self-inactivating (SIN) lentiviral vector. Unlike viral promoters, the enhancer-less A2UCOE gave rise to populations of cells that expressed a reporter transgene at a highly reproducible level. The efficiency of expression per vector genome was also markedly increased in vivo compared with vectors incorporating either spleen focus-forming virus (SFFV) or cytomegalovirus (CMV) promoters, suggesting a relative resistance to silencing. Furthermore, an A2UCOE-IL2RG vector fully restored the IL-2 signaling pathway within IL2RG-deficient human cells in vitro and successfully rescued the X-linked severe combined immunodeficiency (SCID-X1) phenotype in a mouse model of this disease. These data indicate that the A2UCOE displays highly reliable transcriptional activity within a lentiviral vector, largely overcoming insertion-site position effects and giving rise to therapeutically relevant levels of gene expression. These properties are achieved in the absence of classic enhancer activity and therefore may confer a high safety profile.
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影响因子：
5.1
作者：
Kraunus, J;Schaumann, DHS;Baum, C
通讯作者：
Baum, C
影响因子：
20.3
作者：
Modlich, U;Kustikova, OS;Baum, C
通讯作者：
Baum, C
影响因子：
11.4
作者：
COLLIS, P;ANTONIOU, M;GROSVELD, F
通讯作者：
GROSVELD, F
DOI：
10.1073/pnas.91.7.2567
发表时间：
1994-03-29
影响因子：
11.1
作者：
CHALLITA, PM;KOHN, DB
通讯作者：
KOHN, DB
影响因子：
5.4
作者：
HOEBEN, RC;MIGCHIELSEN, AAJ;VANDEREB, AJ
通讯作者：
VANDEREB, AJ