Journal
GENESIS
Volume 45, Issue 5, Pages 275-281Publisher
WILEY-LISS
DOI: 10.1002/dvg.20300
Keywords
HPRT minigene; selection cassette; gene targeting; alternative splicing; chimeric transcript
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Funding
- NCI NIH HHS [R01 CA76317-05A1, T32 CA86800-03] Funding Source: Medline
- NIA NIH HHS [P01 AG17242] Funding Source: Medline
- NIEHS NIH HHS [U01 ES11044] Funding Source: Medline
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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 mini-gene'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.
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