TY - JOUR
T1 - Optimized gene transduction in human lung organoids
T2 - A high-efficiency method for advanced research applications
AU - Khateeb, Jasmin
AU - Liang, Jady
AU - Li, Yuchong
AU - Thanabalasingam, Thenuka
AU - Khang, Julie
AU - Jerkic, Mirjana
AU - Pellecchia, Giovanna
AU - Thiruv, Bhooma
AU - Chen, Ya Wen
AU - Rotstein, Ori
AU - Slutsky, Arthur S.
AU - Zhang, Haibo
N1 - Publisher Copyright:
© The Author(s) 2025.
PY - 2025/12
Y1 - 2025/12
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85217850617&partnerID=8YFLogxK
U2 - 10.1038/s42003-025-07461-w
DO - 10.1038/s42003-025-07461-w
M3 - Article
C2 - 39900972
AN - SCOPUS:85217850617
SN - 2399-3642
VL - 8
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 164
ER -