MICER targeting vectors for manipulating the mouse genome.

The mouse has become an important model for understanding human development, physiology and disease because of its genetic and biological similarity to humans. Desired mouse mutants with precise genetic alterations can now be generated through gene targeting in mouse embryonic stem cells. The rate-limiting factor in a gene-targeting experiment is the time needed for cloning to construct targeting vectors. The establishment of the Mutagenic Insertion and Chromosome Engineering Resource has made available targeting vectors for the insertional mutagenesis of a large number of mouse genes as well as for chromosome engineering throughout the mouse genome. This unique resource has enriched the repertoire of the genetic reagents for targeted manipulation of the mouse genome.

Neuronal differentiation of NTE-deficient embryonic stem cells.

Organophosphates induce neurological disorders. One of the enzymes inhibited by these compounds is neuropathy target esterase (NTE). In vitro, inhibition of NTE activity by organophosphates is correlated with inhibition of neurite initiation and reduction of neurite length, supporting the hypothesis that organophosphate-induced neurological disorders are caused by inhibition of NTE activity. However, there is no direct evidence for the involvement of NTE in organophosphate-induced impairment of neurites in vitro. To examine the role of NTE, we have generated NTE-deficient mouse embryonic stem cells. These cells can differentiate into neuron-like cells. Although NTE-deficient cells exhibited a delay in neurite initiation in vitro, both the proportion of neuron-like cells which initiated neurites and the elongation of these neurites occurred at the normal rate. These results demonstrate that NTE activity is not required for neurite initiation or elongation per se, but is essential for the optimal rate of neurite initiation.