Fig. 1 ∣. The steps of lentivirus packaging and transduction.
The protocol has seven parts: (1) preparation of lentivirus vector with the CRISPR–Cas9 targeting gene of interest; (2) lentivirus packaging; (3) lentivirus transduction of HIEs; (4) HIE recovery and antibiotic selection; (5) single-cell cloning; (6) sequencing verification of single-cell clones; (7) using KO lines to evaluate host gene function in viral replication or other biological processes.