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Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides
β‐thalassaemia is a prevalent hereditary haematological disease caused by mutations in the human haemoglobin β (HBB) gene. Among them, the HBB IVS2‐654 (C > T) mutation, which is in the intron, creates an aberrant splicing site. Bone marrow transplantation for curing β‐thalassaemia is limited due...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6850948/ https://www.ncbi.nlm.nih.gov/pubmed/31631510 http://dx.doi.org/10.1111/jcmm.14669 |
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author | Xiong, Zeyu Xie, Yingjun Yang, Yi Xue, Yanting Wang, Ding Lin, Shouheng Chen, Diyu Lu, Dian He, Lina Song, Bing Yang, Yinghong Sun, Xiaofang |
author_facet | Xiong, Zeyu Xie, Yingjun Yang, Yi Xue, Yanting Wang, Ding Lin, Shouheng Chen, Diyu Lu, Dian He, Lina Song, Bing Yang, Yinghong Sun, Xiaofang |
author_sort | Xiong, Zeyu |
collection | PubMed |
description | β‐thalassaemia is a prevalent hereditary haematological disease caused by mutations in the human haemoglobin β (HBB) gene. Among them, the HBB IVS2‐654 (C > T) mutation, which is in the intron, creates an aberrant splicing site. Bone marrow transplantation for curing β‐thalassaemia is limited due to the lack of matched donors. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9), as a widely used tool for gene editing, is able to target specific sequence and create double‐strand break (DSB), which can be combined with the single‐stranded oligodeoxynucleotide (ssODN) to correct mutations. In this study, according to two different strategies, the HBB IVS2‐654 mutation was seamlessly corrected in iPSCs by CRISPR/Cas9 system and ssODN. To reduce the occurrence of secondary cleavage, a more efficient strategy was adopted. The corrected iPSCs kept pluripotency and genome stability. Moreover, they could differentiate normally. Through CRISPR/Cas9 system and ssODN, our study provides improved strategies for gene correction of β‐Thalassaemia, and the expression of the HBB gene can be restored, which can be used for gene therapy in the future. |
format | Online Article Text |
id | pubmed-6850948 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-68509482019-12-01 Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides Xiong, Zeyu Xie, Yingjun Yang, Yi Xue, Yanting Wang, Ding Lin, Shouheng Chen, Diyu Lu, Dian He, Lina Song, Bing Yang, Yinghong Sun, Xiaofang J Cell Mol Med Original Articles β‐thalassaemia is a prevalent hereditary haematological disease caused by mutations in the human haemoglobin β (HBB) gene. Among them, the HBB IVS2‐654 (C > T) mutation, which is in the intron, creates an aberrant splicing site. Bone marrow transplantation for curing β‐thalassaemia is limited due to the lack of matched donors. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9), as a widely used tool for gene editing, is able to target specific sequence and create double‐strand break (DSB), which can be combined with the single‐stranded oligodeoxynucleotide (ssODN) to correct mutations. In this study, according to two different strategies, the HBB IVS2‐654 mutation was seamlessly corrected in iPSCs by CRISPR/Cas9 system and ssODN. To reduce the occurrence of secondary cleavage, a more efficient strategy was adopted. The corrected iPSCs kept pluripotency and genome stability. Moreover, they could differentiate normally. Through CRISPR/Cas9 system and ssODN, our study provides improved strategies for gene correction of β‐Thalassaemia, and the expression of the HBB gene can be restored, which can be used for gene therapy in the future. John Wiley and Sons Inc. 2019-10-21 2019-12 /pmc/articles/PMC6850948/ /pubmed/31631510 http://dx.doi.org/10.1111/jcmm.14669 Text en © 2019 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Xiong, Zeyu Xie, Yingjun Yang, Yi Xue, Yanting Wang, Ding Lin, Shouheng Chen, Diyu Lu, Dian He, Lina Song, Bing Yang, Yinghong Sun, Xiaofang Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title | Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title_full | Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title_fullStr | Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title_full_unstemmed | Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title_short | Efficient gene correction of an aberrant splice site in β‐thalassaemia iPSCs by CRISPR/Cas9 and single‐strand oligodeoxynucleotides |
title_sort | efficient gene correction of an aberrant splice site in β‐thalassaemia ipscs by crispr/cas9 and single‐strand oligodeoxynucleotides |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6850948/ https://www.ncbi.nlm.nih.gov/pubmed/31631510 http://dx.doi.org/10.1111/jcmm.14669 |
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