|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 4. XCR2 MO blocks endogenous RNA splicing. (A) Genomic structure
including exon I (1), exon II (2), and exon III (3) of XCR2. The
translation initiation codon (ATG) and a potential termination codon (TAA) are
indicated. The splice-site-targeted antisense morpholino oligonucleotide XCR2
MO (MO) was designed against the sequence at the boundary of exon I/intron I.
Abnormal splicing detected in XCR2 MO-injected embryos is shown by the gray
line (v). The spliced variant is generated by a cryptic splice donor (csd)
site 642 bp downstream from the end of exon I. (B) Sequence comparison
of cDNA from mature mRNA and the aberrant splicing variant. Amino acid
sequences are also indicated. The variant generated by abnormal splicing has a
termination codon (correspondent to TAA in
Fig. 4A) right after the end of
exon I. Arrowhead indicates the correct exon I/exon II splice junction. The
target sequence of XCR2 MO is underlined. (C) RT-PCR analyses of
XCR2 mRNA in XCR2 MO-injected embryos. Embryos were injected with 10,
20 and 50 ng of XCR2 MO, or 20 and 50 ng of Standard Control MO (SC MO), and
harvested for RT-PCR at stage 15, 25 and 35. The primer set was designed to
span the first intron to detect splicing of the endogenous XCR2
pre-mRNA. In XCR2 MO-injected embryos, the mature mRNA (m) was decreased and
an abnormal splicing variant (v) was detected. At stage 35, the mature
XCR2 mRNA was detectable at low levels in embryos injected with
lowest dose (10 ng) of XCR2 MO.
|
