Pcdh ef1 mcs pgk copgfp lentiviral expression vector

SLFN11 cDNA was amplified using the forward primer (5′- ATCGGATCC GCGGCCAACATGGAGGCAAATCAGTGC-3′) and the reverse primer with the sequence for the Flag tag (5′-ATTGTCGACGCGGCCCTACTTATCGT CGTCAT CCTTGTAATCATGGCCACCCCACGGAA-3′) and cloned into the pCDH-EF1-MCS-(PGK-copGFP) lentiviral expression vector (System Biosciences) using the In-Fusion HD cloning kit (Clontech). The lentiviral SLFN11-expressing vector and the pPACKH1 lentivector packaging plasmids were cotransfected into 293TN cells (System Biosciences) and the viral particles were collected to infect K562 cells with TransduxTM (System Biosciences). The SLFN11-expressing cells with GFP signal were sorted using a Fluorescence Activated Cell Sorter (FACS).

SLFN11 cDNA was amplified using the forward primer (5′-ATCGGATCCGCGGCCAACATGGAGGCAAATCAGTGC-3′) and the reverse primer with the sequence for the Flag tag (5′-ATTGTCGACGCGGCCCTACTTATCGTCGTCATCCTTGTAATCATGGCCACCCCACGGAA-3′) and cloned into pCDH-EF1-MCS-PGK-copGFP lentiviral expression vector (System Biosciences) by In-Fusion HD cloning kit (Clontech). SLFN11 mutant (E669Q) was generated using the primer (5′-CGTCATTGACGAAGCTCACAATTTCCGTACTGAAGATG-3′) and QuikChange II XL site-directed mutagenesis kit (Agilent Technologies), and the mutation was validated by sequence analysis. The lentiviral SLFN11-expressing vector and the pPACKH1 lentivector packaging plasmids were co-transfected into 293TN cells (System Biosciences) and the viral particles were collected to infect K562 cells with Transdux^™ (System Biosciences). The SLFN11-expressing cells with GFP expression were sorted using a Fluorescence Activated Cell Sorter (FACS).