S/MAR sequence confers long-term mitotic stability on non-integrating lentiviral vector episomes without selection.
S/MAR sequence confers long-term mitotic stability on non-integrating lentiviral vector episomes without selection.
复制标题
S/MAR序列赋予了不选择的非整合慢病毒载体插图的长期有丝分裂稳定性。
DOI：
10.1093/nar/gku082
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Kurre P
中科院分区：
文献类型：
--
作者：
Verghese SC;Goloviznina NA;Skinner AM;Lipps HJ;Kurre P
插入癌基因激活和异常剪接已被证明是逆转录病毒干细胞基因治疗的主要挫折。整合酶缺陷人类免疫缺陷病毒1衍生载体提供了一种潜在的更安全的方法，但它们的环状基因组在细胞分裂过程中迅速丢失。在这里，我们描述了一种新的慢病毒载体（LV），它结合了人类ß-干扰素支架/基质相关区域序列，为外核LTR环的长期有丝分裂维持提供了复制的起源。由此产生的“锚定”非整合慢病毒载体（aniLV）实现了与整合载体相当的初始转导率，随后在一部分细胞中逐步建立长期的外源性表达。对anilv转导的单细胞衍生克隆进行的分析显示，在没有选择压力的情况下维持了100轮细胞分裂，从发作体中持续表达转基因，并为长期维持发作体提供了分子证据。为了评估aniLV在原代细胞中的表现，我们转导了谱系缺失的小鼠造血祖细胞，观察了移植到条件宿主后克隆祖细胞菌落中GFP的表达和外周血白细胞嵌合现象。总的来说，我们的研究表明，支架/基质相关区域元件可以作为非整合慢载体片段的分子锚点，在体外和体内通过连续的细胞分裂和祖细胞分化提供持续的基因表达。
Insertional oncogene activation and aberrant splicing have proved to be major setbacks for retroviral stem cell gene therapy. Integrase-deficient human immunodeficiency virus-1-derived vectors provide a potentially safer approach, but their circular genomes are rapidly lost during cell division. Here we describe a novel lentiviral vector (LV) that incorporates human ß-interferon scaffold/matrix-associated region sequences to provide an origin of replication for long-term mitotic maintenance of the episomal LTR circles. The resulting ‘anchoring’ non-integrating lentiviral vector (aniLV) achieved initial transduction rates comparable with integrating vector followed by progressive establishment of long-term episomal expression in a subset of cells. Analysis of aniLV-transduced single cell-derived clones maintained without selective pressure for >100 rounds of cell division showed sustained transgene expression from episomes and provided molecular evidence for long-term episome maintenance. To evaluate aniLV performance in primary cells, we transduced lineage-depleted murine hematopoietic progenitor cells, observing GFP expression in clonogenic progenitor colonies and peripheral blood leukocyte chimerism following transplantation into conditioned hosts. In aggregate, our studies suggest that scaffold/matrix-associated region elements can serve as molecular anchors for non-integrating lentivector episomes, providing sustained gene expression through successive rounds of cell division and progenitor differentiation in vitro and in vivo.
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影响因子：
12.4
作者：
Modlich, Ute;Navarro, Susana;Baum, Christopher
通讯作者：
Baum, Christopher
影响因子：
12.4
作者：
Kurre, P;Anandakumar, P;Kiem, HP
通讯作者：
Kiem, HP
影响因子：
5.4
作者：
Agarwal, M;Austin, TW;Plavec, I
通讯作者：
Plavec, I
DOI：
10.1186/1479-0556-9-1
发表时间：
2011-01-04
期刊：
Genetic vaccines and therapy
影响因子：
--
作者：
Grandchamp, Nicolas;Henriot, Dorothee;Sarkis, Chamsy
通讯作者：
Sarkis, Chamsy
影响因子：
5.3
作者：
Leight, ER;Sugden, B
通讯作者：
Sugden, B