title: HPRT minigene generates chimeric transcripts as a by-product of gene targeting.
abstract: The HPRT minigene is a selection cassette used for gene targeting in mouse embryonic stem (ES) cells and, it is unique since selection may be applied for its presence and absence. This minigene has two exon clusters separated by a small intron and splicing sequences. We find these exon clusters splice into exons from the target gene forming two different classes of chimeric transcripts. The first class is expressed by the endogenous promoter and includes upstream target gene exons spliced into minigene exons 3-8. The second class is expressed by the minigene's PGK promoter and includes minigene exons 1-2 spliced into downstream target gene exons. These chimeric transcripts may produce chimeric proteins that could influence phenotype. Therefore, we have designed two floxed HPRT minigenes that permit removal of either the 5' half of the minigene or the entire minigene via Cre-mediated recombination.
authors: Department of Molecular Medicine, The Institute of Biotechnology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245-3207, USA.
authors: Valerie B
authors: VB
authors: Holcomb
authors: Tae Moon
authors: TM
authors: Kim
authors: Lavinia C
authors: LC
authors: Dumitrache
authors: Sheng-Mei
authors: SM
authors: Ma
authors: Ming-Jiu
authors: MJ
authors: Chen
authors: Paul
authors: P
authors: Hasty
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002678:Chimera / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism; D005091:Exons; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D051379:Mice; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D007041:Hypoxanthine Phosphoribosyltransferase