Raman multiplexers for alternative gene splicing.
Nonfluorescent labels were used for an array-format multiplex detection of alternative splice junctions of breast cancer susceptibility gene 1 (BRCA1) by surface-enhanced Raman scattering (SERS). A four-plex detection scheme using nonfluorescent labels was demonstrated for DNA sequences specific to four BRCA1 alternative splice variants: Delta(11q) (the last 3309 nt deleted from exon 11), Delta(9, 10) (exon 9 and 10 deleted), Delta(5) (exon 5 deleted), and Delta(5q, 6) (the last 22 nt of exon 5 and the entire exon 6 is deleted). This is the first proof-of-concept study to apply SERS-based detection using nonfluorescent labels to investigate alternative gene splicing. Detection sensitivity of up to 1 fM was demonstrated for the Raman labels chosen to clearly discriminate the splice junctions via specific target identification. The proposed approach has the potential to become a highly sensitive and selective tool for comprehensive alternative splicing profiling of BRCA1 or other genes relevant to specific diseases.
Sun
Lan
L
Department of Agricultural and Biological Engineering and Bindley Bioscience Center, Purdue University, 225 South University Street, West Lafayette, Indiana 47907, USA.
Yu
Chenxu
C
Irudayaraj
Joseph
J
D017398:Alternative Splicing; D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry; D019398:Genes, BRCA1; D006046:Gold / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*
D015342:DNA Probes; D009841:Oligonucleotides; D006046:Gold; D004247:DNA