Using Crispr Cas9 Technique At Mouse Tracheal Cells Progenitor Cells
CRISPR–Cas9 has triggered a revolution in which laboratories around the world are using the technology for innovative applications in biology. CRISPR can efficiently and precisely target, modify, regulate and mark genes in cells and in organisms. To establish a CRISPR genome editing platform, accumulating hands-on experience of applying such technique is necessary. In this experiment, foxj1 has been selected as the target gene as it is essential for ciliated cells formation and its function has been well understood. The whole experimental procedures include: design mfoxj1 sgRNA, sub-cloning lentiCRISPR-Cas9 vector, confirming sgRNA insertion by sequencing, packaging lentiCRISPR-Cas9 capsids, transfecting mouse tracheal epithelial progenitor cells and evaluating the knock-out results. Although the knock-out results are not available now, previous experimental procedures have provided valuable experience, which can facilitate to establish a CRISRP gene editing platform in the laboratory.
Keywords: CRISPR-Cas9 system, mouse foxj1, mTECs, ALI cell culture
Functionality of CRISPR-Cas9
CRISPR-Cas9 genome editing technology has exploded in popularity these years, which has been rapidly adopted by the scientific community around the world. This novel technique: RNA-guided endonuclease Cas9, which originates from the adaptive bacterial immunity system CRISPR…