Table 1 Summary of gene-targeting assays in LIG4−/−POLQ−/− cells.

From: Dual loss of human POLQ and LIG4 abolishes random integration

Target locus Targeting vector No. of cells treated ( × 106) Gene-targeting efficiency
  5′-arm 3′-arm Selection marker*   
HPRT 3.8 kb 5.1 kb PPGK -Neo 40.0 100% (138/138)
HPRT 1.7 kb 2.8 kb 2A-Puro 24.0 100% (59/59)
AAVS1 4.4 kb 3.1 kb PPGK -Neo 4.0 100% (8/8)
PRKDC 2.1 kb 5.1 kb PPGK -Hyg 8.0 100% (3/3)
RAD54B 3.3 kb 4.5 kb PPGK -Hyg 8.0 100% (5/5)
RAD54L 2.9 kb 3.6 kb 2A-Puro 8.0 100% (1/1)
TP53BP1 0.9 kb 1.5 kb IRES-Hyg 8.0 100% (2/2)
  1. PPGK, PGK promoter; 2A, 2A peptide sequence.
  2. *It is noteworthy that although Hygr and Puror (hygromycin- and puromycin-resistance gene, respectively) have already been used to disrupt POLQ, increasing drug concentrations allowed us to reuse these genes as gene-targeting selection markers.