Table 1.

Repair of a genomic DSB in the absence or presence of a homologous donor

Donor and cell lineaElectroporationbTotal no. of HAT-resistant coloniesFrequency of HAT-resistant colonies (10−5)c
Construct usedNo.
No donor
 Sce-3I-SceI421570.03
pCMV-I-SceI106500.65
Mock1300
360-bp donor
 360-1pCMV-I-SceI202,6002.6
Mock500
 360-2pCMV-I-SceI202,0002.0
Mock500
2-kb donor
 26-1pCMV-I-SceI56732.7
Mock51010.4
 26-2pCMV-I-SceI81,1562.9
Mock8580.14
  • a Cell line Sce-3 contains an integrated copy of pTKSce (Fig. 1B), cell lines 360-1 and 360-2 each contain an integrated copy of pTKSce-360 (Fig. 1B), and cell lines 26-1 and 26-2 each contain an integrated copy of pTKSce-26 (Fig. 1B).

  • b Cell line Sce-3 was electroporated with 150 U of I-SceI endonuclease or 5 μg of pCMV-I-SceI per electroporation. Other cell lines were electroporated with 10 μg of pCMV-I-SceI per electroporation. Mock electroporations were with PBS alone.

  • c Total number of HAT-resistant colonies recovered divided by total number of cells electroporated.