University of Colorado Anschutz Medical Campus
Must have completed three years of college
Number of Openings: 1
LINE-1 (Long Interspersed Nuclear Element-1, or L1) is a major dynamic force in the mammalian genome. Retrotransposition deposits the progeny of L1 throughout the genome, sometimes leading to gene disruption, modified expression of adjacent genes, and/or transduction of neighboring DNA. In addition, L1, as interspersed repetitive DNA, provides a substrate for homologous recombination of mispaired sequences, leading to gene duplication, deletion, chromosome translocation and exon shuffling.
Any one of the dynamic events caused by the presence and movement of L1 in the human genome can lead to disease including cancer. Thus, it is extremely important to understand the details of the intermediates involved in retrotransposition and the mechanisms used to control their expression and movement in vivo.
The immediate goal of this work will be to determine the molecular basis for translational control of the production of the two L1-encoded proteins.
We have identified several cellular proteins that interact with L1 RNA regions that are hypothesized to control the production of the second L1 protein in L1’s dicistronic mRNA. Experiments will use siRNA to knockdown four of these proteins and assess the effect of this knockdown on translation using reporter constructs.
Additionally, retrotransposition assays will be used to determine the effect of knockdown of these proteins on L1 retrotransposition in cultured cells. Methods used: mammalian cell culture, plasmid DNA isolation and analysis, cell transfection, luciferase assay, eGFP assay by flow cytometry.