Lent-On-Plus Lentiviral vectors for conditional expression in human stem cells

被引：15

作者：

Benabdellah, Karim ^{[1
,2
]}

Munoz, Pilar ^{[1
,3
]}

Cobo, Marien ^{[1
]}

Gutierrez-Guerrero, Alejandra ^{[1
]}

Sanchez-Hernandez, Sabina ^{[1
]}

Garcia-Perez, Angelica ^{[1
,4
]}

Anderson, Per ^{[1
]}

Belen Carrillo-Galvez, Ana ^{[1
]}

Toscano, Miguel G. ^{[1
,5
]}

Martin, Francisco ^{[1
,2
]}

机构：

[1] Pfizer Univ Granada Andalusian Reg Goverment, Gen Med Dept, GENYO, Ctr Genom & Oncol Res, Parque Tecnol Ciencias Salud,Ave Ilustrac 114, Granada 18016, Spain

[2] Pfizer Univ Granada Andalusian Reg Gov, Ctr Genom & Oncol Res, LentiStem Biotech, GENYO, Parque Tecnol Ciencias Salud,Ave Ilustrac 114, Granada 18016, Spain

[3] UCL, Inst Child Hlth, 30 Guilford Street, London WC1N 1EH, England

[4] Max Delbruck Ctr Mol Med, Berlin, Germany

[5] Amarna Therapeut S L, Marie Curie Bldg,C Leonardo da Vinci 18a, Seville 41092, Spain

来源：

SCIENTIFIC REPORTS | 2016年 / 6卷

关键词：

SCAFFOLD ATTACHMENT REGION; GENE-EXPRESSION; TRANSGENE EXPRESSION; IN-VITRO; HEMATOPOIETIC DIFFERENTIATION; RETROVIRAL EXPRESSION; RNA INTERFERENCE; CHS4; INSULATOR; EFFICIENT; METHYLATION;

D O I：

10.1038/srep37289

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

Conditional transgene expression in human stem cells has been difficult to achieve due to the low efficiency of existing delivery methods, the strong silencing of the transgenes and the toxicity of the regulators. Most of the existing technologies are based on stem cells clones expressing appropriate levels of tTA or rtTA transactivators (based on the TetR-VP16 chimeras). In the present study, we aim the generation of Tet-On all-in-one lentiviral vectors (LVs) that tightly regulate transgene expression in human stem cells using the original TetR repressor. By using appropriate promoter combinations and shielding the LVs with the Is2 insulator, we have constructed the Lent-On-Plus Tet-On system that achieved efficient transgene regulation in human multipotent and pluripotent stem cells. The generation of inducible stem cell lines with the Lent-ON-Plus LVs did not require selection or cloning, and transgene regulation was maintained after long-term cultured and upon differentiation toward different lineages. To our knowledge, Lent-On-Plus is the first all-in-one vector system that tightly regulates transgene expression in bulk populations of human pluripotent stem cells and its progeny.

引用

页数：17

共 77 条

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