Floxin, a resource for genetically engineering mouse ESCs

Veena Singla, Julie Hunkapiller, Nicole Santos, Allen D Seol, Andrew R Norman, Paul Wakenight, William C Skarnes, Jeremy F Reiter
<span title="2009-12-06">2009</span> <i title="Springer Nature"> <a target="_blank" rel="noopener" href="https://fatcat.wiki/container/l6cw2tyvgrhytbvantrj2os37y" style="color: black;">Nature Methods</a> </i> &nbsp;
We describe a method for the highly efficient and precise targeted modification of gene trap loci in mouse embryonic stem cells (ESCs). Through the Floxin method, gene trap mutations are reverted and new DNA sequences inserted using Cre recombinase and a shuttle vector, pFloxin. Floxin technology is applicable to the existing collection of 24,149 compatible gene trap cell lines, which should enable the high-throughput modification of many genes in mouse ESCs. Genetic modification of mice and of
more &raquo; ... embryonic stem cells (ESCs) is an important source of insight into the functions of vertebrate genes. Current methods commonly used for genetically modifying ESCs are transgene insertion and homologous recombination. Transgene insertion involves integration at a random location in the genome. Homologous recombination can create targeted mutations, but only about 1.5% of ESC clones, on average, correctly integrate the construct and different targeting constructs must be created for each gene. A complementary approach to generating loss-of-function mutations in ESCs is gene trapping1-3. Below, we describe the gene trap vectors pGTLxf and pGTLxr, which allow post-insertional modification of the trapped locus using an accompanying technology called Users may view, print, copy, download and text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:
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