In this report we demonstrate primary non-random X chromosome inactivation
following targeted mutagenesis of a region immediately upstream of Xist pro
moter P-1. In heterozygous animals there is a preferential inactivation of
the targeted X chromosome in 80-90% of cells, The phenotype correlates with
inappropriate activation of Xist in a proportion of the mutant XY embryoni
c stem cells. Strand-specific analysis revealed increased sense transcripti
on initiating upstream of Xist promoter P-1. There was, however, no discern
ible effect on transcription from the antisense Tsix gene, We demonstrate t
hat the in vitro and in vivo phenotypes are specifically attributable to th
e presence of a PGKneo cassette at the targeted locus. These findings are d
iscussed in the context of understanding mechanisms of Xist gene regulation
in X inactivation.