LiPS-A3S, a human genomic site for robust expression of inserted transgenes
Transgenesis of human pluripotent stem cells (hPSCs) can enable and empower a variety of studies in stem cell research, including lineage tracing and functional genetics studies. While in recent years much progress has been made in the development of tools for gene targeting, little attention has be...
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2016-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253117301191 |
| _version_ | 1818505773631668224 |
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| author | Andriana G Kotini Michel Sadelain Eirini P Papapetrou |
| author_facet | Andriana G Kotini Michel Sadelain Eirini P Papapetrou |
| author_sort | Andriana G Kotini |
| collection | DOAJ |
| description | Transgenesis of human pluripotent stem cells (hPSCs) can enable and empower a variety of studies in stem cell research, including lineage tracing and functional genetics studies. While in recent years much progress has been made in the development of tools for gene targeting, little attention has been given to the identification of sites in the human genome where transgenes can be inserted and reliably expressed. In order to find human genomic sites capable of supporting long-term and high-level transgene expression in hPSCs, we performed a lentiviral screen in human induced pluripotent stem cells (iPSCs). We isolated 40 iPSC clones each harboring a single vector copy and characterized the level of transgene expression afforded by each unique integration site. We selected one clone, LiPS-A3 with an integration site in chromosome 15 maintaining robust expression without silencing and demonstrate that different transgenes can be inserted therein rapidly and efficiently through recombinase-mediated cassette exchange (RMCE). The LiPS-A3 line can greatly facilitate the insertion of reporter and other genes in hPSCs. Targeting transgenes in the LiPS-A3S genomic locus can find broad applications in stem cell research and possibly cell and gene therapy. |
| first_indexed | 2024-12-10T21:55:29Z |
| format | Article |
| id | doaj.art-496d5314f73a4d38ba447852dfdb9b53 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-10T21:55:29Z |
| publishDate | 2016-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-496d5314f73a4d38ba447852dfdb9b532022-12-22T01:32:04ZengElsevierMolecular Therapy: Nucleic Acids2162-25312016-01-015C10.1038/mtna.2016.99LiPS-A3S, a human genomic site for robust expression of inserted transgenesAndriana G Kotini0Michel Sadelain1Eirini P Papapetrou2Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USACenter for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York, New York, USADepartment of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USATransgenesis of human pluripotent stem cells (hPSCs) can enable and empower a variety of studies in stem cell research, including lineage tracing and functional genetics studies. While in recent years much progress has been made in the development of tools for gene targeting, little attention has been given to the identification of sites in the human genome where transgenes can be inserted and reliably expressed. In order to find human genomic sites capable of supporting long-term and high-level transgene expression in hPSCs, we performed a lentiviral screen in human induced pluripotent stem cells (iPSCs). We isolated 40 iPSC clones each harboring a single vector copy and characterized the level of transgene expression afforded by each unique integration site. We selected one clone, LiPS-A3 with an integration site in chromosome 15 maintaining robust expression without silencing and demonstrate that different transgenes can be inserted therein rapidly and efficiently through recombinase-mediated cassette exchange (RMCE). The LiPS-A3 line can greatly facilitate the insertion of reporter and other genes in hPSCs. Targeting transgenes in the LiPS-A3S genomic locus can find broad applications in stem cell research and possibly cell and gene therapy.http://www.sciencedirect.com/science/article/pii/S2162253117301191 |
| spellingShingle | Andriana G Kotini Michel Sadelain Eirini P Papapetrou LiPS-A3S, a human genomic site for robust expression of inserted transgenes Molecular Therapy: Nucleic Acids |
| title | LiPS-A3S, a human genomic site for robust expression of inserted transgenes |
| title_full | LiPS-A3S, a human genomic site for robust expression of inserted transgenes |
| title_fullStr | LiPS-A3S, a human genomic site for robust expression of inserted transgenes |
| title_full_unstemmed | LiPS-A3S, a human genomic site for robust expression of inserted transgenes |
| title_short | LiPS-A3S, a human genomic site for robust expression of inserted transgenes |
| title_sort | lips a3s a human genomic site for robust expression of inserted transgenes |
| url | http://www.sciencedirect.com/science/article/pii/S2162253117301191 |
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