کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10533573 | 961884 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Stemless self-quenching reporter molecules identify target sequences in mRNA
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
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چکیده انگلیسی
The design of oligonucleotides for gene silencing requires a rational method for identifying hybridization-accessible sequences within the target RNA. To this end, we have developed stem-loop self-quenching reporter molecules (SQRMs) as probes for such sequence. SQRMs have a 5â² fluorophore, a quenching moiety on the 3â² end, an intervening sequence that forms an â¼5-basepaired stem, and a loop sequence of â¼20-30 bases. We have previously described a mapping strategy employing SQRMs to locate stem-loop structures in the target mRNA molecule. We now show that the original design constraint of a basepaired stem is not needed, either in vitro or in vivo. We propose that stemless probes possess sufficient signal-to-noise for use in vivo and that this ratio is an indication of hybridization of the probe to its target. Data showing that these SQRMs can specifically target and reduce c-Myb protein synthesis and can be used for real-time in vivo assays are presented.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 347, Issue 1, 1 December 2005, Pages 77-88
Journal: Analytical Biochemistry - Volume 347, Issue 1, 1 December 2005, Pages 77-88
نویسندگان
Lida K. Gifford, David Jordan, Vikram Pattanayak, Kathy Vernovsky, Bao T. Do, Alan M. Gewirtz, Ponzy Lu,