Target accessibility and signal specificity in live-cell detection of BMP-4 mRNA using molecular beacons | Zendy

Won Jong Rhee | Zendy; Philip J. Santangelo | Zendy; Hanjoong Jo | Zendy; Gang Bao | Zendy

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Target accessibility and signal specificity in live-cell detection of BMP-4 mRNA using molecular beacons

Author(s) -

Won Jong Rhee,

Philip J. Santangelo,

Hanjoong Jo,

Gang Bao

Publication year - 2008

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gkn039

Subject(s) - molecular beacon , biology , gene knockdown , messenger rna , microbiology and biotechnology , rna , in situ hybridization , computational biology , genetics , oligonucleotide , gene

The ability to visualize mRNA in single living cells and monitor in real-time the changes of mRNA level and localization can provide unprecedented opportunities for biological and disease studies. However, the mRNA detection specificity and sensitivity are critically dependent on the selection of target sequences and their accessibility. We carried out an extensive study of the target accessibility of BMP-4 mRNA using 10 different designs of molecular beacons (MBs), and identified the optimal beacon design. Specifically, for MB design 1 and 8 (MB1 and MB8), the fluorescent intensities from BMP-4 mRNA correlated well with the GFP signal after upregulating BMP-4 and co-expressing GFP using adenovirus, and the knockdown of BMP-4 mRNA using siRNA significantly reduced the beacon signals, demonstrating detection specificity. The beacon specificity was further confirmed using blocking RNA and in situ

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