One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes
Neural cell fate is determined by a tightly controlled transcription regulatory network during development. The ability to manipulate the expression of multiple transcription factors simultaneously is required to delineate the complex picture of neural cell development. Because of the limited carryi...
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doaj-b0bc94377a754274b8595762a6a51caa2020-11-24T22:09:18ZengElsevierMolecular Therapy: Nucleic Acids2162-25312017-09-0186476One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple GenesShenglan Li0Anqi Zhang1Haipeng Xue2Dali Li3Ying Liu4The Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX 77030, USAThe Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Central Laboratory, Liaocheng People’s Hospital, 252000 Shandong, ChinaThe Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX 77030, USAThe Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX 77030, USAThe Vivian L. Smith Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Corresponding author: Ying Liu, Department of Neurosurgery, The University of Texas Health Science Center at Houston, 1825 Pressler Street, SRB 630G, Houston, TX 77030, USA.Neural cell fate is determined by a tightly controlled transcription regulatory network during development. The ability to manipulate the expression of multiple transcription factors simultaneously is required to delineate the complex picture of neural cell development. Because of the limited carrying capacity of the commonly used viral vectors, such as lentiviral or retroviral vectors, it is often challenging to perform perturbation experiments on multiple transcription factors. Here we have developed a piggyBac (PB) transposon-based CRISPR activation (CRISPRa) all-in-one system, which allows for simultaneous and stable endogenous transactivation of multiple transcription factors and long non-coding RNAs. As a proof of principle, we showed that the PB-CRISPRa system could accelerate the differentiation of human induced pluripotent stem cells into neurons and astrocytes by triggering endogenous expression of different sets of transcription factors. The PB-CRISPRa system has the potential to become a convenient and robust tool in neuroscience, which can meet the needs of a variety of in vitro and in vivo gain-of-function applications. Keywords: CRISPR, gene activation, human induced pluripotent stem cells, piggyBac, transposonhttp://www.sciencedirect.com/science/article/pii/S2162253117301920 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shenglan Li Anqi Zhang Haipeng Xue Dali Li Ying Liu |
spellingShingle |
Shenglan Li Anqi Zhang Haipeng Xue Dali Li Ying Liu One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes Molecular Therapy: Nucleic Acids |
author_facet |
Shenglan Li Anqi Zhang Haipeng Xue Dali Li Ying Liu |
author_sort |
Shenglan Li |
title |
One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes |
title_short |
One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes |
title_full |
One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes |
title_fullStr |
One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes |
title_full_unstemmed |
One-Step piggyBac Transposon-Based CRISPR/Cas9 Activation of Multiple Genes |
title_sort |
one-step piggybac transposon-based crispr/cas9 activation of multiple genes |
publisher |
Elsevier |
series |
Molecular Therapy: Nucleic Acids |
issn |
2162-2531 |
publishDate |
2017-09-01 |
description |
Neural cell fate is determined by a tightly controlled transcription regulatory network during development. The ability to manipulate the expression of multiple transcription factors simultaneously is required to delineate the complex picture of neural cell development. Because of the limited carrying capacity of the commonly used viral vectors, such as lentiviral or retroviral vectors, it is often challenging to perform perturbation experiments on multiple transcription factors. Here we have developed a piggyBac (PB) transposon-based CRISPR activation (CRISPRa) all-in-one system, which allows for simultaneous and stable endogenous transactivation of multiple transcription factors and long non-coding RNAs. As a proof of principle, we showed that the PB-CRISPRa system could accelerate the differentiation of human induced pluripotent stem cells into neurons and astrocytes by triggering endogenous expression of different sets of transcription factors. The PB-CRISPRa system has the potential to become a convenient and robust tool in neuroscience, which can meet the needs of a variety of in vitro and in vivo gain-of-function applications. Keywords: CRISPR, gene activation, human induced pluripotent stem cells, piggyBac, transposon |
url |
http://www.sciencedirect.com/science/article/pii/S2162253117301920 |
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