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Cells containing integrated construct (via gene targeting or random integration) were then selected in medium containing Puromycin.
Lanes 1 and 3: hESCs with random integration of the targeting construct.
Ribosomal DNA (rDNA) repeat sequences are overrepresented during random integration of rAAV.
The homologous recombination events were confirmed by Southern blotting, further indicating the lack of any random integration (Fig. 1D).
Such random integration may result in alteration of gene expression or gene disruption by unpredictable position effects.
In most cell types, the frequency of homologous recombination is markedly lower than the frequency of random integration.
As shown in Fig. 1D, two clones were identified to have precise HR at both arms and no random integration.
For the selection against random integration in mouse ES cells, it contains the HSV TK gene.
HGn39D was generated by random integration of an enhancer-trap transgene [28].
The transgenic technique is associated with the problem of random integration [31], [34].
Plasmid transfection may lead to random integration into the genome at low frequency.
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