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. Author manuscript; available in PMC: 2014 Aug 11.
Published in final edited form as: Mol Ther. 2007 Oct 23;16(1):187–193. doi: 10.1038/sj.mt.6300339

Figure 4. Analysis of exon skipping after gene targeting at COL1A2 exon 4.

Figure 4

(a) Sodium dodecyl sulfate polyacrylamide gel electrophoresis separation of [3H]-proline-labeled collagen peptides from OIMSC12 clones targeted at the mutant (TM) or wild-type (TW) COL1A2 allele. Light bands indicated by asterisks (*) are α2(I) chains produced from the targeted alleles. (b) Reverse transcriptase-PCR (RT-PCR) of transcripts of the 5’-untranslated region (UTR)-exon 8 region of COL1A2 from wild-type (WT) and gene-targeted clones, labeled as in a. Numbers at right correspond to PCR products from (1) a heteroduplex product composed of WT and exon 4-skipped complementary DNA (cDNA); (2) the normal, full-length messenger RNA (mRNA); (3) mRNA missing exon 4; and (4) mRNA missing exons 3 and 4. (c) Reamplification products of excised cDNA species 1–4 from b. (d) Schematic drawing of mRNA with lines indicating alternative-spliced forms detected, and PCR primers used for RT-PCR (arrows). 5’-UTR (black box) and coding exons (open boxes) are shown.