Selection of keratinocytes transduced with the multidrug resistance gene in an in vitro skin model presents a strategy for enhancing gene expression in vivo.
Selection of keratinocytes transduced with the multidrug resistance gene in an in vitro skin model presents a strategy for enhancing gene expression in vivo.
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在体外皮肤模型中选择转导多药耐药基因的角质形成细胞提出了增强体内基因表达的策略。
DOI：
10.1089/10430349950016546
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发表时间：
1999
影响因子：
4.2
通讯作者：
J. Vogel
中科院分区：
文献类型：
--
作者：
W. Pfützner;U. Hengge;Mohamed A. Joari;R. Foster;J. Vogel
在基因治疗研究中，很难在相当大比例的细胞中实现长时间的高水平基因表达。增强表达的一种解决方案是选择在双顺反子载体中表达所需基因和连接的选择标记基因的细胞。作为潜在的靶组织，皮肤易于安全地局部应用选择剂，这可能导致高百分比的角质形成细胞中显著的基因表达。为了测试这种方法的可行性，开发了皮筏养殖模型。人角质形成细胞转导的多药耐药（MDR）基因，赋予各种细胞生长抑制和抗有丝分裂化合物，如秋水仙碱的阻力。当在无细胞真皮上生长时，用各种剂量的秋水仙碱（10-50 ng/ml）处理转导的角质形成细胞。秋水仙素处理增加角质形成细胞表达MDR的百分比，几乎100%的筏培养物中，明显地，角质形成细胞在秋水仙素处理的，MDR转导的筏培养物中能够正常增殖并形成分层的分化表皮。该模型表明，局部选择MDR表达的角质形成细胞在体内应该是可行的，而不妨碍皮肤的生物完整性。因此，局部选择导致与抗性基因相关的所需基因的表达增强，为皮肤基因治疗带来了未来的希望。
In gene therapy studies, achieving prolonged, high-level gene expression in a significant percentage of cells has been difficult. One solution to enhance expression would be to select for cells expressing both the desired gene and a linked selectable marker gene in a bicistronic vector. As a potential target tissue, the skin is easily accessible for safe topical application of a selecting agent that could lead to significant gene expression in a high percentage of keratinocytes. To test the feasibility of such an approach, a skin raft culture model was developed. Human keratinocytes were transduced with the multidrug resistance (MDR) gene, which confers resistance to a variety of cytostatic and antimitotic compounds, such as colchicine. While growing on acellular dermis, transduced keratinocytes were treated with various doses of colchicine (10-50 ng/ml). Colchicine treatment increased the percentage of keratinocytes expressing MDR to almost 100% in raft cultures, Significantly, keratinocytes in colchicine-treated, MDR-transduced raft cultures were able to proliferate normally and form a stratified, differentiated epidermis. This model suggests that topical selection for MDR-expressing keratinocytes in vivo should be feasible without hampering the biologic integrity of skin. Thus, topical selection leading to enhanced expression of a desired gene, linked to a resistance gene, holds future promise for skin gene therapy.
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遗传性表皮疾病的基因纠正。
DOI：
10.1089/104303400750035816
发表时间：
2000
期刊：
Human gene therapy.
影响因子：
--
作者：
Khavari,PA
通讯作者：
Khavari,PA
器官培养中逆转录病毒介导的猪角质形成细胞转导。
DOI：
10.1046/j.1523-1747.1998.00298.x
发表时间：
1998
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
Ghazizadeh,S;Harrington,R;Garfield,J;Taichman,LB
通讯作者：
Taichman,LB
基因皮肤病：基因治疗的候选疾病。
DOI：
10.1089/104303400750035807
发表时间：
2000
期刊：
Human gene therapy.
影响因子：
--
作者：
Uitto,J;Pulkkinen,L
通讯作者：
Pulkkinen,L
表皮：基因治疗的一个有吸引力的靶组织。
DOI：
10.1111/1523-1747.ep12399070
发表时间：
1994
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
Greenhalgh,DA;Rothnagel,JA;Roop,DR
通讯作者：
Roop,DR
DOI：
10.1073/pnas.95.8.4356
发表时间：
1998-04-14
影响因子：
11.1
作者：
Kolodka, TM;Garlick, JA;Taichman, LB
通讯作者：
Taichman, LB