Journal
TRANSGENIC RESEARCH
Volume 18, Issue 3, Pages 483-489Publisher
SPRINGER
DOI: 10.1007/s11248-008-9237-9
Keywords
Protein prenylation; Farnesylation; Prelamin A; HDJ-2; Knockout mice
Categories
Funding
- Ellison Medical Foundation Senior Scholar Award
- National Institutes of Health (NIH) [HL76839, CA099506, AR050200, HL086683]
- Dr. Mariano Barbacid (Spain)
- American Heart Association, Western States Affiliate
- NATIONAL CANCER INSTITUTE [R01CA099506] Funding Source: NIH RePORTER
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL089781, R01HL076839, R01HL086683] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES [R01AR050200] Funding Source: NIH RePORTER
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A common strategy for conditional knockout alleles is to flox (flank with loxP sites) a 5' exon within the target gene. Typically, the floxed exon does not contain a unit number of codons so that the Cre-mediated recombination event yields a frameshift and a null allele. Documenting recombination within the genomic DNA is often regarded as sufficient proof of a frameshift, and the analysis of transcripts is neglected. We evaluated a previously reported conditional knockout allele for the beta-subunit of protein farnesyltransferase. The recombination event in that allele-the excision of exon 3-was predicted to yield a frameshift. However, following the excision of exon 3, exon 4 was skipped by the mRNA splicing machinery, and the predominant transcript from the mutant allele lacked exon 3 and exon 4 sequences. The a dagger exon 3-4 transcript does not contain a frameshift but rather is predicted to encode a protein with a short in-frame deletion. This represents a significant concern when studying an enzyme, since an enzyme with partial function could lead to erroneous conclusions. With thousands of new conditional knockout alleles under construction within mouse mutagenesis consortiums, the protein farnesyltransferase allele holds an important lesson-to characterize knockout alleles at both the DNA and RNA levels.
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