PMID- 36484412
OWN - NLM
STAT- MEDLINE
DCOM- 20230117
LR  - 20230117
IS  - 1364-5528 (Electronic)
IS  - 0003-2654 (Linking)
VI  - 148
IP  - 2
DP  - 2023 Jan 16
TI  - A highly specific and flexible detection assay using collaborated actions of 
      DNA-processing enzymes for identifying multiple gene expression signatures in 
      breast cancer.
PG  - 316-327
LID - 10.1039/d2an01672a [doi]
AB  - Most nucleic acid biosensors employ nucleic acid-processing enzymes to bind, 
      degrade, splice, synthesize, and modify nucleic acids. Utilizing their unique 
      substrate preference, binding mode, and catalytic activity is of great importance 
      in designing nucleic acid biosensors. Combination with DNA-processing enzymes 
      enables them to transform into a new generation of molecular diagnostics tools 
      with enhanced selectivity and sensitivity and reduced reaction time. Here, we 
      report an isothermal amplification strategy by coemploying a structure-specific 
      endonuclease (flap endonuclease 1, FEN1) and a strand-displacing DNA polymerase 
      (Bst DNA polymerase) to detect long RNA targets. This approach couples the 
      FEN1-driven invasive cleavage reaction with toehold-mediated rolling circle 
      amplification (iFEN-tRCA), enabling the highly selective and rapid detection of 
      long RNA targets and offering a detection limit below 10 pM within 1 h. We used 
      two targets, such as human epidermal growth factor receptor 2 (HER2, encoded by 
      ERBB2) and dopamine- and cyclic AMP-regulated phosphoprotein (DARPP, encoded by 
      PPP1R1B), associated with prognosis or response to anticancer therapy. We 
      demonstrated the feasibility and quantitative capability of the iFEN-tRCA assay 
      by assessing the expression of two RNA transcripts (ERBB2 and PPP1R1B) with total 
      RNA extracts purified from human breast cancer cells. Therefore, we envision that 
      the developed assay will provide a suitable prognostic and diagnostic tool for 
      identifying appropriate patients for HER2-targeted therapy and predicting the 
      clinical outcome and occurrence of metastasis relapse in breast cancer.
FAU - Kim, Dain
AU  - Kim D
AD  - Department of Bioengineering & Nano-bioengineering, Research Center for Bio 
      Materials and Process Development, Incheon National University, Incheon 22012, 
      Republic of Korea. e.kim@inu.ac.kr.
FAU - Lee, Jiyoung
AU  - Lee J
AD  - Division of Bioengineering, Incheon National University, Incheon 22012, Republic 
      of Korea.
FAU - Han, Jueun
AU  - Han J
AD  - Department of Chemistry, Incheon National University, Incheon 22012, Republic of 
      Korea.
FAU - Lim, Jaewoo
AU  - Lim J
AD  - BioNanotechnology Research Center, Korea Research Institute of Bioscience and 
      Biotechnology (KRIBB), Daejeon 34141, Republic of Korea. eklim1112@kribb.re.kr.
FAU - Lim, Eun-Kyung
AU  - Lim EK
AUID- ORCID: 0000-0003-2793-3700
AD  - BioNanotechnology Research Center, Korea Research Institute of Bioscience and 
      Biotechnology (KRIBB), Daejeon 34141, Republic of Korea. eklim1112@kribb.re.kr.
AD  - Department of Nanobiotechnology, KRIBB School of Biotechnology, UST, Daejeon 
      34113, Republic of Korea.
AD  - School of Pharmacy, Sungkyunkwan University, Suwon 16419, Republic of Korea.
FAU - Kim, Eunjung
AU  - Kim E
AUID- ORCID: 0000-0001-9579-9414
AD  - Department of Bioengineering & Nano-bioengineering, Research Center for Bio 
      Materials and Process Development, Incheon National University, Incheon 22012, 
      Republic of Korea. e.kim@inu.ac.kr.
AD  - Division of Bioengineering, Incheon National University, Incheon 22012, Republic 
      of Korea.
LA  - eng
PT  - Journal Article
DEP - 20230116
PL  - England
TA  - Analyst
JT  - The Analyst
JID - 0372652
RN  - 9007-49-2 (DNA)
RN  - EC 2.7.7.7 (DNA-Directed DNA Polymerase)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Humans
MH  - Female
MH  - *Breast Neoplasms/diagnosis/genetics
MH  - Transcriptome
MH  - Nucleic Acid Amplification Techniques
MH  - DNA/genetics/metabolism
MH  - DNA-Directed DNA Polymerase/chemistry
MH  - RNA
EDAT- 2022/12/10 06:00
MHDA- 2023/01/18 06:00
CRDT- 2022/12/09 06:43
PHST- 2022/12/10 06:00 [pubmed]
PHST- 2023/01/18 06:00 [medline]
PHST- 2022/12/09 06:43 [entrez]
AID - 10.1039/d2an01672a [doi]
PST - epublish
SO  - Analyst. 2023 Jan 16;148(2):316-327. doi: 10.1039/d2an01672a.