Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon.
Compact Bidirectional Promoters for Dual-Gene Expression in a Sleeping Beauty Transposon.
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DOI：
10.3390/ijms21239256
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发表时间：
2020-12-04
影响因子：
5.6
通讯作者：
McLellan AD
中科院分区：
文献类型：
--
作者：
He K;Rad SMAH;Poudel A;McLellan AD
启动子的选择是基因治疗中转基因表达的重要考虑因素。多个基因的表达需要核糖体进入或跳跃位点，或使用多个启动子。由两个分开的、不同的启动子组成的启动子系统可以显著增加预期用于基因治疗的基因盒的大小。然而，另一种方法是使用单一、紧凑、双向启动子。我们鉴定了RPBSA合成启动子的强且稳定的双向活性，所述RPBSA合成启动子由人Rpl 13 a启动子的片段以及另外的内含子/外显子结构组成。基于Rpl 13 a的启动子驱动荧光蛋白的长期双向活性。对于EF 1-α和LMP 2/TAP 1启动子获得了类似的结果。然而，在慢病毒载体中，趋异双向系统未能产生足够的滴度以翻译成用于双重嵌合抗原受体（CAR）表达的表达系统。尽管双向启动子显示出在睡美人转座子系统中驱动短RNA的优异适用性，但它们在需要更长和更复杂RNA的慢病毒应用（例如双CAR盒）中的可能用途是有限的。
Promoter choice is an essential consideration for transgene expression in gene therapy. The expression of multiple genes requires ribosomal entry or skip sites, or the use of multiple promoters. Promoter systems comprised of two separate, divergent promoters may significantly increase the size of genetic cassettes intended for use in gene therapy. However, an alternative approach is to use a single, compact, bidirectional promoter. We identified strong and stable bidirectional activity of the RPBSA synthetic promoter comprised of a fragment of the human Rpl13a promoter, together with additional intron/exon structures. The Rpl13a-based promoter drove long-term bidirectional activity of fluorescent proteins. Similar results were obtained for the EF1-α and LMP2/TAP1 promoters. However, in a lentiviral vector, the divergent bidirectional systems failed to produce sufficient titres to translate into an expression system for dual chimeric antigen receptor (CAR) expression. Although bidirectional promoters show excellent applicability to drive short RNA in Sleeping Beauty transposon systems, their possible use in the lentiviral applications requiring longer and more complex RNA, such as dual-CAR cassettes, is limited.
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影响因子：
4.5
作者：
Collins PJ;Kobayashi Y;Nguyen L;Trinklein ND;Myers RM
通讯作者：
Myers RM
影响因子：
4.4
作者：
Jangid RK;Kelkar A;Muley VY;Galande S
通讯作者：
Galande S
影响因子：
14.9
作者：
Anno YN;Myslinski E;Ngondo-Mbongo RP;Krol A;Poch O;Lecompte O;Carbon P
通讯作者：
Carbon P
影响因子：
7
作者：
Trinklein, ND;Aldred, SF;Myers, RM
通讯作者：
Myers, RM
影响因子：
3.7
作者：
Rad, Ali Hosseini S. M.;Poudel, Aarati;McLellan, Alexander D.
通讯作者：
McLellan, Alexander D.