title: SINE Retrotransposition: Evaluation of Alu Activity and Recovery of De Novo Inserts.
abstract: Mobile element activity is of great interest due to its impact on genomes. However, the types of mobile elements that inhabit any given genome are remarkably varied. Among the different varieties of mobile elements, the Short Interspersed Elements (SINEs) populate many genomes, including many mammalian species. Although SINEs are parasites of Long Interspersed Elements (LINEs), SINEs have been highly successful in both the primate and rodent genomes. When comparing copy numbers in mammals, SINEs have been vastly more successful than other nonautonomous elements, such as the retropseudogenes and SVA. Interestingly, in the human genome the copy number of Alu (a primate SINE) outnumbers LINE-1 (L1) copies 2 to 1. Estimates suggest that the retrotransposition rate for Alu is tenfold higher than LINE-1 with about 1 insert in every twenty births. Furthermore, Alu-induced mutagenesis is responsible for the majority of the documented instances of human retroelement insertion-induced disease. However, little is known on what contributes to these observed differences between SINEs and LINEs. The development of an assay to monitor SINE retrotransposition in culture has become an important tool for the elucidation of some of these differences. In this chapter, we present details of the SINE retrotransposition assay and the recovery of de novo inserts. We also focus on the nuances that are unique to the SINE assay.
authors.affiliation: Department of Epidemiology, Tulane Cancer Center, SL-66, Tulane University Health Sciences Center, 1430 Tulane Ave., New Orleans, LA, 70112, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Ade
authors.affiliation: Department of Epidemiology, Tulane Cancer Center, SL-66, Tulane University Health Sciences Center, 1430 Tulane Ave., New Orleans, LA, 70112, USA. aengel@tulane.edu.
authors.forename: Astrid M
authors.initials: AM
authors.lastname: Roy-Engel
mesh_terms: D020087:Alu Elements; D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D016366:Open Reading Frames; D020086:Short Interspersed Nucleotide Elements; D014162:Transfection
keywords: Alu; ORF2 protein; RNA polymerase III; Retrotransposition; SINE