Protocol for odorant-binding protein 2A gene knockdown by dual siRNA transfection in a three-dimensional human epidermal equivalent model
Summary: Gene knockdown by small interfering RNA (siRNA) transfection is widely used to investigate gene function. Here, we present a protocol for the knockdown of the odorant-binding protein 2A (OBP2A) gene in a three-dimensional human epidermal equivalent model (3DE-model). We describe steps for g...
Saved in:
| Main Author: | |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725000322 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Summary: | Summary: Gene knockdown by small interfering RNA (siRNA) transfection is widely used to investigate gene function. Here, we present a protocol for the knockdown of the odorant-binding protein 2A (OBP2A) gene in a three-dimensional human epidermal equivalent model (3DE-model). We describe steps for growing human epidermal keratinocytes (normal human epithelial keratinocytes [NHEKs]) for 3DE-model construction in monolayer culture. We then detail procedures for transfecting the cells with siRNA, followed by a second siRNA transfection during 3DE-model construction. The efficiency of gene knockdown is verified by qPCR and ELISA.For complete details on the use and execution of this protocol, please refer to Nakanishi et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
|---|---|
| ISSN: | 2666-1667 |