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Generating a novel cell line with a codon optimized LINE1 element at a single locus
Author(s) -
Russell Brittany,
Morales Maria,
DeFreece Cecily
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.lb196
Subject(s) - endonuclease , retrotransposon , biology , genetics , genome , locus (genetics) , start codon , dna , messenger rna , gene , transposable element
Long interspersed element‐1 (LINE1) is the only currently active autonomous retroelement within the human genome. LINE1 replicates itself through a process known as retrotransposition that requires the endonuclease and reverse transcriptase activities of the LINE1 protein ORF2. Further studies characterizing LINE1 endonuclease function demonstrated that LINE1 endonuclease induces formation of double‐stranded breaks (DSBs). Only a small fraction of LINE1‐induced DSBs results in a retrotransposition event, suggesting that the damage from the LINE1 endonuclease activity may be greater than previously considered. While LINE1 mutagenic insertion events have been associated with diseases such as breast and colon cancer as well as muscular dystrophy, the full effect of LINE1 endonuclease upon stability of the genome is undetermined. By inserting a codon‐optimized LINE1 element into a cell line at a unique genomic locus using the FRT system, we will investigate and track the effect of the endonuclease on the genome. Codon optimization of LINE1 will increase expression of the element and will aid in detection of novel retrotransposition events in the presences and absence of drugs. Here we are presenting the cloning of the FRT vector containing the codon‐optimized LINE1 element, the first step of project. Support or Funding Information The work presented is supported in part by monies from P20GM103424, TL4GM118968 and 5RL5GM118966‐03 as well as the Louisiana Cancer Research Consortium.