Single point mutation detection in living cancer cells by far-red emitting PNA–FIT probes

Abstract

Peptide nucleic acid bis-quinoline conjugates are reported as attractive far-red emitting probes that detect mutated mRNA in living cells at SNP resolution.

Enhanced SNP-sensing using DNA-templated reactions through confined hybridization of minimal substrates (CHOMS)

“…12,13 The need to achieve more costeffective, faster analysis and higher single nucleotide resolution continues to drive technological developments. 14 Recent examples include amendments to existing technologies such as molecular beacons, 15,16 melting analysis, 17,18 environmentally sensitive uorescent nucleobases, [19][20][21][22][23][24] and strand displacement probes 25,26 or new technologies such as polymerase-amplied release of ATP (POLARA) 27 or graphene-based biosensors for real-time kinetic monitoring of hybridization. 28 The analysis of SNVs requires technologies with the highest nucleotide resolution to ascertain the polymorphism or variation.…”

Monitoring Dicer‐Mediated miRNA‐21 Maturation and Ago2 Loading by a Dual‐Colour FIT PNA Probe Set

“…The 11‐nt PNA strand 1 is complementary to the U38‐A48 segment of pre‐miR21. The red emitting quinoline blue (QB) dye was introduced at various positions (Table S1). This “QB‐walk” revealed FIT PNA 1 as a probe providing a 14‐fold enhancement of fluorescence upon addition of pre‐miR21 (Figure A, Table ).…”

Nucleic‐Acid Structures as Intracellular Probes for Live Cells

“…Furthermore, FIT probes offer fluorescence enhancement, reported to be as high as 195‐fold upon addition of complementary target . Much of the work in live cell imaging using FIT probes has relied upon the use of peptide nucleic acids (PNA) due to their biostability and both rapid and strong hybridization to complementary ONTs . Kam et al, for example, used PNA FIT probes for detection of KRAS mRNA inside live cells .…”

“…[154] Much of the work in live cell imaging using FIT probes has relied upon the use of peptide nucleic acids (PNA) due to their biostability and both rapid and strong hybridization to complementary ONTs. [79][80][81]155,156] Kam et al, for example, used PNA FIT probes for detection of KRAS mRNA inside live cells. [79] They showed that FIT probes are sensitive to single nucleotide mismatches directly adjacent to the intercalator, such that single nucleotide polymorphisms (SNPs) in the KRAS gene could be discriminated.…”