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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.
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BMC Research Notes

ISSN: 1756-0500

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