التفاصيل البيبلوغرافية
| العنوان: |
Transcription Activator-Like Effector Nuclease (TALEN)-Mediated CLYBL Targeting Enables Enhanced Transgene Expression and One-Step Generation of Dual Reporter Human Induced Pluripotent Stem Cell (iPSC) and Neural Stem Cell (NSC) Lines. |
| المؤلفون: |
Cerbini, Trevor, Funahashi, Ray, Luo, Yongquan, Liu, Chengyu, Park, Kyeyoon, Rao, Mahendra, Malik, Nasir, Zou, Jizhong |
| المصدر: |
PLoS ONE; Jan2015, Vol. 10 Issue 1, p1-18, 18p |
| مصطلحات موضوعية: |
NUCLEASES, GENETIC transcription, TRANSGENE expression, PLURIPOTENT stem cells, NEURAL stem cells |
| مستخلص: |
Targeted genome engineering to robustly express transgenes is an essential methodology for stem cell-based research and therapy. Although designer nucleases have been used to drastically enhance gene editing efficiency, targeted addition and stable expression of transgenes to date is limited at single gene/locus and mostly PPP1R12C/AAVS1 in human stem cells. Here we constructed transcription activator-like effector nucleases (TALENs) targeting the safe-harbor like gene CLYBL to mediate reporter gene integration at 38%–58% efficiency, and used both AAVS1-TALENs and CLYBL-TALENs to simultaneously knock-in multiple reporter genes at dual safe-harbor loci in human induced pluripotent stem cells (iPSCs) and neural stem cells (NSCs). The CLYBL-TALEN engineered cell lines maintained robust reporter expression during self-renewal and differentiation, and revealed that CLYBL targeting resulted in stronger transgene expression and less perturbation on local gene expression than PPP1R12C/AAVS1. TALEN-mediated CLYBL engineering provides improved transgene expression and options for multiple genetic modification in human stem cells. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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