Generation of SFTPC-mCherry knock-in reporter human embryonic stem cell line, WAe001-A-2H, using CRISPR/Cas9-based gene targeting

Hui Sun; Qian Li; Tao Xu; Wei Zhang; Jiaqi Sun; Huisheng Liu

doi:10.1016/j.scr.2024.103597

Generation of SFTPC-mCherry knock-in reporter human embryonic stem cell line, WAe001-A-2H, using CRISPR/Cas9-based gene targeting

Stem Cell Res. 2024 Dec:81:103597. doi: 10.1016/j.scr.2024.103597. Epub 2024 Oct 28.

Authors

Hui Sun¹, Qian Li², Tao Xu¹, Wei Zhang³, Jiaqi Sun⁴, Huisheng Liu⁵

Affiliations

¹ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou National Laboratory, Guangzhou 510005, China.
² Guangzhou National Laboratory, Guangzhou 510005, China.
³ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou National Laboratory, Guangzhou 510005, China; Bioland Laboratory, Guangzhou 510005, China. Electronic address: [email protected].
⁴ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou National Laboratory, Guangzhou 510005, China; Bioland Laboratory, Guangzhou 510005, China. Electronic address: [email protected].
⁵ School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Guangzhou National Laboratory, Guangzhou 510005, China; Bioland Laboratory, Guangzhou 510005, China. Electronic address: [email protected].

PMID: 39476617
DOI: 10.1016/j.scr.2024.103597

Abstract

The SFTPC gene is responsible for the production of the pulmonary surfactant protein C (SPC), a highly hydrophobic molecule that plays a crucial role in maintaining lung integrity through its influence on the synthesis of alveolar surfactant proteins. In this study, we harnessed the CRISPR/Cas9 system for precise genome editing to create a modified H1 human embryonic stem cell (hESC) line, incorporating the SFTPC-mCherry reporter construct. Therefore, the engineered SFTPC-mCherry knock-in (KI) hESC line can serve as an effective tool for tracking the expression patterns of the SFTPC gene as alveolar type 2 cells differentiate from hESCs.

MeSH terms

CRISPR-Cas Systems*
Cell Line
Gene Editing / methods
Gene Knock-In Techniques* / methods
Gene Targeting / methods
Genes, Reporter
Human Embryonic Stem Cells* / cytology
Human Embryonic Stem Cells* / metabolism
Humans
Pulmonary Surfactant-Associated Protein C* / genetics
Pulmonary Surfactant-Associated Protein C* / metabolism

Substances

Pulmonary Surfactant-Associated Protein C
SFTPC protein, human