Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells

TET1-mediated active DNA demethylation is required for endogenous retrovirus (ERV) enhancer activation during human ES differentiation into definitive endoderm (DE) cells. Here we present a protocol for siRNA-mediated TET1 knockdown during this process to decipher TET1’s role in ERV activation and D...

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Detalles Bibliográficos
Autores principales: Liufu, Zhongqi, Liu, Yujian, He, Jiangping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10371804/
https://www.ncbi.nlm.nih.gov/pubmed/37467109
http://dx.doi.org/10.1016/j.xpro.2023.102455
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author Liufu, Zhongqi
Liu, Yujian
He, Jiangping
author_facet Liufu, Zhongqi
Liu, Yujian
He, Jiangping
author_sort Liufu, Zhongqi
collection PubMed
description TET1-mediated active DNA demethylation is required for endogenous retrovirus (ERV) enhancer activation during human ES differentiation into definitive endoderm (DE) cells. Here we present a protocol for siRNA-mediated TET1 knockdown during this process to decipher TET1’s role in ERV activation and DE differentiation. We describe steps for inducing ES into DE cells. We then detail steps for knocking down TET1 during differentiation and for examining the effects of TET1 knockdown on LTR6B methylation, cell morphology, and gene expression. For complete details on the use and execution of this protocol, please refer to Wu et al. (2022).(1)
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spelling pubmed-103718042023-07-28 Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells Liufu, Zhongqi Liu, Yujian He, Jiangping STAR Protoc Protocol TET1-mediated active DNA demethylation is required for endogenous retrovirus (ERV) enhancer activation during human ES differentiation into definitive endoderm (DE) cells. Here we present a protocol for siRNA-mediated TET1 knockdown during this process to decipher TET1’s role in ERV activation and DE differentiation. We describe steps for inducing ES into DE cells. We then detail steps for knocking down TET1 during differentiation and for examining the effects of TET1 knockdown on LTR6B methylation, cell morphology, and gene expression. For complete details on the use and execution of this protocol, please refer to Wu et al. (2022).(1) Elsevier 2023-07-18 /pmc/articles/PMC10371804/ /pubmed/37467109 http://dx.doi.org/10.1016/j.xpro.2023.102455 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Liufu, Zhongqi
Liu, Yujian
He, Jiangping
Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title_full Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title_fullStr Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title_full_unstemmed Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title_short Protocol for siRNA-mediated TET1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
title_sort protocol for sirna-mediated tet1 knockdown during differentiation of human embryonic stem cells into definitive endoderm cells
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10371804/
https://www.ncbi.nlm.nih.gov/pubmed/37467109
http://dx.doi.org/10.1016/j.xpro.2023.102455
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