Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system
Human induced pluripotent stem (hiPS) cells are a powerful tool for biomedical research. The ability to create patient-specifc pluripotent cells and their subsequent differentiation into any somatic cell type makes hiPS cells a valuable object for creating in vitro models of human diseases, screening...
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| Format: | Article |
| Language: | English |
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Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2019-01-01
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| Series: | Вавиловский журнал генетики и селекции |
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| Online Access: | https://vavilov.elpub.ru/jour/article/view/1805 |
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| _version_ | 1832575159168401408 |
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| author | М. М. Gridina |
| author_facet | М. М. Gridina |
| author_sort | М. М. Gridina |
| collection | DOAJ |
| description | Human induced pluripotent stem (hiPS) cells are a powerful tool for biomedical research. The ability to create patient-specifc pluripotent cells and their subsequent differentiation into any somatic cell type makes hiPS cells a valuable object for creating in vitro models of human diseases, screening drugs and a future source of cells for regenerative medicine. To realize entirely a potential of hiPScells, effective and precise methods for their genome editing are needed. The CRISPR/Cas9 system is the most widely used method for introducing site-specifc double-stranded breaks into DNA. It allows genes of interest to be knocked out with high efciency. However, knock-in into the target site of the genome is a much more difcult task. Moreover, many researchers have noted a low efciency of introducing target constructs into the hiPS cells’ genome. In this review, I attempt to describe the currently known information regarding the matter of increasing efciency of targeted insertions into hiPS cells’ genome. Here I will describe the most effective strategies for designing the donor template for homology-directed repair, methods to manipulate the double-strand break repair pathways introduced by a nuclease, including control of CRISPR/Cas9 delivery time. A low survival rate of hiPS cells following genome editing experiments is another difculty on the way towards successful knock-in, and here several highly effective approaches addressing it are proposed. Finally, I describe the most promising strategies, one-step reprogramming and genome editing, which allows gene-modifed integration-free hiPS cells to be efciently generated directly from somatic cells. |
| format | Article |
| id | doaj-art-5e9f8192928344b9bbc79bd0aa00355f |
| institution | Kabale University |
| issn | 2500-3259 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders |
| record_format | Article |
| series | Вавиловский журнал генетики и селекции |
| spelling | doaj-art-5e9f8192928344b9bbc79bd0aa00355f2025-02-01T09:58:06ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592019-01-012281026103210.18699/VJ18.446853Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 systemМ. М. Gridina0Institute of Cytology and Genetics, SB RASHuman induced pluripotent stem (hiPS) cells are a powerful tool for biomedical research. The ability to create patient-specifc pluripotent cells and their subsequent differentiation into any somatic cell type makes hiPS cells a valuable object for creating in vitro models of human diseases, screening drugs and a future source of cells for regenerative medicine. To realize entirely a potential of hiPScells, effective and precise methods for their genome editing are needed. The CRISPR/Cas9 system is the most widely used method for introducing site-specifc double-stranded breaks into DNA. It allows genes of interest to be knocked out with high efciency. However, knock-in into the target site of the genome is a much more difcult task. Moreover, many researchers have noted a low efciency of introducing target constructs into the hiPS cells’ genome. In this review, I attempt to describe the currently known information regarding the matter of increasing efciency of targeted insertions into hiPS cells’ genome. Here I will describe the most effective strategies for designing the donor template for homology-directed repair, methods to manipulate the double-strand break repair pathways introduced by a nuclease, including control of CRISPR/Cas9 delivery time. A low survival rate of hiPS cells following genome editing experiments is another difculty on the way towards successful knock-in, and here several highly effective approaches addressing it are proposed. Finally, I describe the most promising strategies, one-step reprogramming and genome editing, which allows gene-modifed integration-free hiPS cells to be efciently generated directly from somatic cells.https://vavilov.elpub.ru/jour/article/view/1805human induced pluripotent stem cellscrispr/cas9 systemgenome editingknock-in efciency |
| spellingShingle | М. М. Gridina Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system Вавиловский журнал генетики и селекции human induced pluripotent stem cells crispr/cas9 system genome editing knock-in efciency |
| title | Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system |
| title_full | Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system |
| title_fullStr | Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system |
| title_full_unstemmed | Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system |
| title_short | Improvement of the knock-in effciency in the genome of human induced pluripotent stem cells using the CRISPR/Cas9 system |
| title_sort | improvement of the knock in effciency in the genome of human induced pluripotent stem cells using the crispr cas9 system |
| topic | human induced pluripotent stem cells crispr/cas9 system genome editing knock-in efciency |
| url | https://vavilov.elpub.ru/jour/article/view/1805 |
| work_keys_str_mv | AT mmgridina improvementoftheknockineffciencyinthegenomeofhumaninducedpluripotentstemcellsusingthecrisprcas9system |