Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications
Abstract Human induced pluripotent stem cell (iPSC)-derived lung organoids, engineered to carry targeted genes, offer a robust platform for investigating mechanistic insights in lung research. Although lentiviral vectors (LVVs) are highly effective for stable expression due to their integrative prop...
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| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-02-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07461-w |
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| _version_ | 1823861760020447232 |
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| author | Jasmin Khateeb Jady Liang Yuchong Li Thenuka Thanabalasingam Julie Khang Mirjana Jerkic Giovanna Pellecchia Bhooma Thiruv Ya-Wen Chen Ori Rotstein Arthur S. Slutsky Haibo Zhang |
| author_facet | Jasmin Khateeb Jady Liang Yuchong Li Thenuka Thanabalasingam Julie Khang Mirjana Jerkic Giovanna Pellecchia Bhooma Thiruv Ya-Wen Chen Ori Rotstein Arthur S. Slutsky Haibo Zhang |
| author_sort | Jasmin Khateeb |
| collection | DOAJ |
| description | Abstract Human induced pluripotent stem cell (iPSC)-derived lung organoids, engineered to carry targeted genes, offer a robust platform for investigating mechanistic insights in lung research. Although lentiviral vectors (LVVs) are highly effective for stable expression due to their integrative properties, achieving efficient transduction in human iPSC-derived lung organoids poses a significant technical challenge, likely due to the complex structure of these organoids. In this study, we optimized a method to enhance LVV transduction efficiency by physically disrupting the organoids to increase surface area, followed by spinoculation to apply shear force during cell dissociation. This approach, combined with the use of an optimized culture medium, significantly improved transduction efficiency. The success of this method was validated at both the gene and protein levels using single-cell RNA sequencing (scRNA-seq) and various cellular and molecular assays. Our optimized transduction protocol may provide a valuable tool for investigating specific cellular and molecular mechanisms in development and disease models using human iPSCs-derived lung organoids. |
| format | Article |
| id | doaj-art-ce9bec4740e9424f8e4892543cca779d |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-ce9bec4740e9424f8e4892543cca779d2025-02-09T12:50:49ZengNature PortfolioCommunications Biology2399-36422025-02-018111010.1038/s42003-025-07461-wOptimized gene transduction in human lung organoids: A high-efficiency method for advanced research applicationsJasmin Khateeb0Jady Liang1Yuchong Li2Thenuka Thanabalasingam3Julie Khang4Mirjana Jerkic5Giovanna Pellecchia6Bhooma Thiruv7Ya-Wen Chen8Ori Rotstein9Arthur S. Slutsky10Haibo Zhang11Keenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoGenetics and Genome Biology, The Hospital for Sick ChildrenGenetics and Genome Biology, The Hospital for Sick ChildrenBlack Family Stem Cell Institute, Department of Cell, Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York cityKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoKeenan Research Centre for Biomedical Science, St. Michael’s Hospital, Unity Health TorontoAbstract Human induced pluripotent stem cell (iPSC)-derived lung organoids, engineered to carry targeted genes, offer a robust platform for investigating mechanistic insights in lung research. Although lentiviral vectors (LVVs) are highly effective for stable expression due to their integrative properties, achieving efficient transduction in human iPSC-derived lung organoids poses a significant technical challenge, likely due to the complex structure of these organoids. In this study, we optimized a method to enhance LVV transduction efficiency by physically disrupting the organoids to increase surface area, followed by spinoculation to apply shear force during cell dissociation. This approach, combined with the use of an optimized culture medium, significantly improved transduction efficiency. The success of this method was validated at both the gene and protein levels using single-cell RNA sequencing (scRNA-seq) and various cellular and molecular assays. Our optimized transduction protocol may provide a valuable tool for investigating specific cellular and molecular mechanisms in development and disease models using human iPSCs-derived lung organoids.https://doi.org/10.1038/s42003-025-07461-w |
| spellingShingle | Jasmin Khateeb Jady Liang Yuchong Li Thenuka Thanabalasingam Julie Khang Mirjana Jerkic Giovanna Pellecchia Bhooma Thiruv Ya-Wen Chen Ori Rotstein Arthur S. Slutsky Haibo Zhang Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications Communications Biology |
| title | Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications |
| title_full | Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications |
| title_fullStr | Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications |
| title_full_unstemmed | Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications |
| title_short | Optimized gene transduction in human lung organoids: A high-efficiency method for advanced research applications |
| title_sort | optimized gene transduction in human lung organoids a high efficiency method for advanced research applications |
| url | https://doi.org/10.1038/s42003-025-07461-w |
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