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    肽核酸研究平臺(tái)

    賽百盛為肽核酸合成提供專業(yè)的解決方案

     

     

  • 什么是肽核酸（PNA）？

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    肽核酸 (Peptide Nucleic Acid，PNA) 是一種人工合成的類似于 DNA 或 RNA的聚合物，最初是由Peter E.Nielsen和他在哥本哈根大學(xué)的同事于1991年合成的 [1]。

    ?

    肽核酸的基本單位是N-(2-氨基乙基)甘氨酸，由通過肽鍵連接形成聚合物 [2]。核酸堿基（ATCG）通過亞甲基羰基鍵而不是通常的脫氧核糖（DNA）或核糖（RNA）連接到主鏈上。與DNA或RNA雙鏈相比，肽核酸與DNA或RNA可以形成更穩(wěn)定的雙鏈結(jié)構(gòu)，并且這一結(jié)構(gòu)代謝穩(wěn)定。

  • 特點(diǎn)

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    高結(jié)合親和力

    電荷中性骨架允許PNA通過Watson-Crick氫鍵與DNA或RNA結(jié)合，其結(jié)合親和力明顯高于帶負(fù)電荷的DNA寡聚體 [3]。

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    高特異性

    PNA序列對(duì)目標(biāo)位點(diǎn)具有極高的特異性。如果有一個(gè)錯(cuò)配，則結(jié)合親和力顯著降低；如果有兩個(gè)錯(cuò)配，則不會(huì)發(fā)生雜交 [4]。

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    高穩(wěn)定性

    PNA可以在DNA或者RNA降解的條件下依然表現(xiàn)出很強(qiáng)的化學(xué)穩(wěn)定性和熱穩(wěn)定性 [5]。

  • PNA 和 DNA 的對(duì)比

    特征

     

    結(jié)合親和力

     

    雜交速率

     

    雜交中的鹽濃度

     

    單堿基錯(cuò)配的ΔTm值

     

    化學(xué)穩(wěn)定性

     

    生物穩(wěn)定性

     

    溶解度

     

    一般探針長(zhǎng)度

    PNA

     

    每個(gè)堿基至少高1°C的Tm值

     

    快100-5000倍

     

    獨(dú)立

     

    <10°C

     

    穩(wěn)定

     

    耐酶

     

    適中（但可通過簡(jiǎn)單修飾改善）

     

    13-18個(gè)堿基

    DNA

     

    -

     

    -

     

    依賴

     

    <15°C

     

    在酸性或堿性條件下不穩(wěn)定

     

    被核酸酶降解

     

    良好

     

    25-30個(gè)堿基

  • 應(yīng)用

    反義失活

    由于PNA具有高結(jié)合親和力，所以可以設(shè)計(jì)PNA靶向mRNA，通過空間干擾以影響其作用 [6]。

    核酸檢測(cè)

    PNA獨(dú)特的雜交特性和代謝穩(wěn)定性使其非常適合于復(fù)雜的生物環(huán)境體系中的核酸檢測(cè) [7]。

    基因編輯

    PNA能夠結(jié)合dsDNA并進(jìn)行基因編輯。經(jīng)過特定修飾以后，PNA介導(dǎo)的基因編輯的脫靶效應(yīng)極弱 [8]。

  • 了解更多

    原則上，熒光原位雜交（FISH）能夠提供個(gè)別染色體端粒長(zhǎng)度的信息。直接標(biāo)記的寡核苷酸探針是這種分析的理想選擇，它們體積小、滲透性能好、單鏈且無需變性，同時(shí)合成過程可控。然而，寡核苷酸雜交用于端粒...
    查看更多...
    具體設(shè)計(jì)規(guī)則: 長(zhǎng)度：合成 PNA 序列一般不超過 18 個(gè)堿基，但不包括接頭、氨基酸和標(biāo)記物。 含有氨基酸：可以把一個(gè)賴氨酸或半胱氨酸聯(lián)在序列的 N 端或 C 端。 嘌呤含量：富含...
    查看更多...
    關(guān)于肽核酸 肽核酸 (Peptide Nucleic Acid，PNA) 是一種人工合成的類似于 DNA 或 RNA 的聚合物。與多肽類似，各種嘌呤和嘧啶堿基通過亞甲基羰基連接到主鏈骨架上。...
    查看更多...

  • 拓展閱讀

    [PNA_Review paper]Tirayut Vilaivan(2018) Beilstein J Org Chem. 14: 253–281

    Fluorogenic PNA probes

    [PNA-FISH] Rajat Gupta, et al.(2018)Cell.173:972-988

    DNA Repair Network Analysis Reveals Shieldin as a Key Regulator of NHEJ and PARP Inhibitor Sensitivity

    [PNA MDx] Hyun Sik Kim, et al.(2018)Pathol Res Pract. pii: S0344-0338(18)30181-X

    Clinical implications of TERT promoter mutation on IDH mutation and MGMT promoter methylation in diffuse gliomas

    [PNA MDx] Jae Young Hur, et al.(2018)Molecular Cancer 17:15

    Extracellular vesicle-derived DNA for performing EGFR genotyping of NSCLC patients

    [PNA probe] Zhu B and Travas-Sejdic J(2018)Analyst. 143(3):687-694

    PNA versus DNA in electrochemical gene sensing based on conducting polymers: study of charge and surface blocking effects on the sensor signal

    [PNA antisense drug] Baek A, et al.(2018)Bioconjug Chem. 29(2):528-537

    Polyethylene Glycol-Engrafted Graphene Oxide as Biocompatible Materials for Peptide Nucleic Acid Delivery into Cells

    [PNA probe] Hu Q, et al.(2018)Biosens Bioelectron. 101:1-6

    Electrochemically mediated in situ growth of electroactive polymers for highly sensitive detection of double-stranded DNA without sequence-preference

    [PNA-FISH] Wang L, et al.(2018)Nanotechnology. 29(28):285602

    Single Molecule Localization Imaging of Telomeres and Centromeres Using Fluorescence in Situ Hybridization and Semiconductor Quantum Dots.

    [PNA probe] Bala A and Górski ?(2018)Bioelectrochemistry. 119:189-195

    Peptide nucleic acid as a selective recognition element for electrochemical determination of Hg2

    [PNA MDx] Jae-Uk Song and Jonghoo Lee(2018)Yonsei Med J. 59: 211-218

    Peptide Nucleic Acid Clamping and Direct Sequencing in the Detection of Oncogenic Alterations in Lung Cancer: Systematic Review and Meta-Analysis

    [PNA-FISH] Lafuente R, et al.(2018)Andrologia. 50(2)

    Sperm telomere length in motile sperm selection techniques: A qFISH approach

    [PNA MDx] Aeri Kim, et al.(2017)J Breast Cancer. 20:150-159

    Mutations of the Epidermal Growth Factor Receptor Gene in Triple-Negative Breast Cancer

    [PNA MDx] Chang Gon Kim, et al.(2017)Oncotarget. 8: 65111–65122

    Detection of activating and acquired resistant mutation in plasma from EGFR-mutated NSCLC patients by peptide nucleic acid(PNA) clamping-assisted fluorescence melting curve analysis

    [PNA MDx] Mi-young Park, et al.(2017) Oncotarget. 8:36331-36338

    Generation of lung cancer cell lines harboring EGFR T790M mutation by CRISPR/Cas9-mediated genome editing

    [PNA-FISH] Ting-Lin B. Yang(2017)Nat Commun. 8:14766

    Mutual reinforcement between telomere capping and canonical Wnt signalling in the intestinal stem cell niche

    [PNA MDx] C. G. Woo, et al.(2017)Annals of Oncology 28:791–797

    Differential protein stability and clinical responses of EML4-ALK fusion variants to various ALK inhibitors in advanced ALK-rearranged non-small cell lung cancer

    [PNA MDx] Ha Young Park, et al.(2016)Thoracic Cancer 7: 639–647

    Quantification of epidermal growth factor receptor (EGFR) mutation may be a predictor of EGFR-tyrosine kinase inhibitor treatment response

    [PNA MDx] Ji-Youn Han, et al.(2016)BMC Cancer 16: 627

    PNA clamping-assisted fluorescence melting curve analysis for detecting EGFR and KRAS mutations in the circulating tumor DNA of patients with advanced non-small cell lung cancer

  • 參考文獻(xiàn)

    1. Nielsen, P. E. , et al. "Sequence-Selective Recognition of DNA by Strand Displacement with a Thymine-Substituted Polyamide." Science?254.5037(1992):1497-1500.
    2. Nielsen, Pe , and G. Haaima . "Peptide nucleic acid (PNA). A DNA mimic with a pseudopeptide backbone."?Chemical Society Reviews?26.2(1997):p. 73-78.
    3. Egholm, M et al. “PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen-bonding rules.”?Nature?vol. 365,6446 (1993): 566-8.?
    4. Nielsen, Peter E. , et al. "Sequence specific inhibition of DNA restriction enzyme cleavage by PNA. "?Nuclc Acids Research?21.2(1993):197.
    5. Uhlmann, Eugen , et al. "PNA: Synthetic Polyamide Nucleic Acids with Unusual Binding Properties."?Angewandte Chemie International Edition?37.20(1998):2796-2823.
    6. Dean, David A. . "Peptide nucleic acids: versatile tools for gene therapy strategies."?Advanced Drug Delivery Reviews?44.2-3(2000):81-95.
    7. Tirayut, Vilaivan . "Fluorogenic PNA probes."?Blstn Journal of Organic Chemistry?14(2018):253-281.
    8. Bahal, Raman et al. “In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery.”?Nature communications?vol. 7 13304. 26 Oct. 2016, doi:10.1038/ncomms13304

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