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Table 2 Effect of the novel negative selection cassette on integration frequency

From: Construction and applications of exon-trapping gene-targeting vectors with a novel strategy for negative selection

Experiment DT-A Random integration frequencya Fold decrease
1 1.0 × 10−4 (1) 25
+ 4.1 × 10−6 (0.04)
2 2.8 × 10−5 (1) 8.5
+ 3.3 × 10−6 (0.12)
3 6.2 × 10−5 (1) 5.6
+ 1.1 × 10−5 (0.18)
  1. The targeting vector pHPRT-SH 2A-EGFP-2A-Puro was I-SceI linearized and transfected into HT1080 cells by using the Nucleofector II system (Experiments 1 and 2) or the MaxCyte devise (Experiment 3). DT-A denotes the presence (+) or absence (−) of the ExTraPANS cassette (SA–IRES–DT-A–polyA sequence) in the targeting vector. The random-integration frequency was determined as described in “Methods”.
  2. aNumbers in parentheses represent relative random-integration frequencies.