Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

Kai Xiong; Yan Zhou; Poul Hyttel; Lars Bolund; Kristine Karla Freude; Yonglun Luo

doi:10.1016/j.scr.2016.10.011

Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

Kai Xiong
, Yan Zhou
, Poul Hyttel
, Lars Bolund
, Kristine Karla Freude^*
, Yonglun Luo

^*Corresponding author for this work

University of Copenhagen
Aarhus University

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.

Original language	English
Journal	Stem Cell Research
Volume	17
Issue number	3
Pages (from-to)	665-669
Number of pages	5
ISSN	1873-5061
DOIs	https://doi.org/10.1016/j.scr.2016.10.011
Publication status	Published - 2016
Externally published	Yes

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2016 SAM iPSCFinal published version, 1.89 MB

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@article{308a750de2654fd695a500cbcf550138,

title = "Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)",

abstract = "Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.",

author = "Kai Xiong and Yan Zhou and Poul Hyttel and Lars Bolund and Freude, \{Kristine Karla\} and Yonglun Luo",

note = "1873-5061/{\textcopyright} 2016 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).",

year = "2016",

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T1 - Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

AU - Xiong, Kai

AU - Zhou, Yan

AU - Hyttel, Poul

AU - Bolund, Lars

AU - Freude, Kristine Karla

AU - Luo, Yonglun

PY - 2016

Y1 - 2016

N2 - Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.

AB - Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4 and C-MYC. We have validated that the reprogramming cassette is silenced in the SAM iPSC clones. Expression of pluripotency genes (OCT4, SOX2, LIN28A, NANOG, GDF3, SSEA4, and TRA-1-60), differentiation potential to all three germ layers, and normal karyotypes are validated. These SAM-iPSCs provide a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes.

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DO - 10.1016/j.scr.2016.10.011

M3 - Journal article

C2 - 27934604

AN - SCOPUS:85020084809

SN - 1873-5061

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SP - 665

EP - 669

JO - Stem Cell Research

JF - Stem Cell Research

IS - 3

ER -

Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

Abstract

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