Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium.
Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium.
复制标题
在气道上皮中提供三肽核酸分子的纳米颗粒。
DOI：
10.1038/ncomms7952
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发表时间：
2015-04-27
影响因子：
16.6
通讯作者：
Egan, Marie E.
中科院分区：
文献类型：
--
作者：
McNeer, Nicole Ali;Anandalingam, Kavitha;Fields, Rachel J.;Caputo, Christina;Kopic, Sascha;Gupta, Anisha;Quijano, Elias;Polikoff, Lee;Kong, Yong;Bahal, Raman;Geibel, John P.;Glazer, Peter M.;Saltzman, W. Mark;Egan, Marie E.
囊性纤维化（CF）是一种致命的遗传性疾病，最常见的原因是囊性纤维化跨膜传导调节因子（CFTR）基因的F508del突变。由于其全身性以及包括体内基因传递和瞬时基因表达在内的挑战，它不容易接受基因治疗。在这里，我们在可生物降解的聚合物纳米颗粒中使用三聚体形成的PNA分子和供体DNA来纠正F508del。我们通过测序和功能性氯化物外排试验确认了修饰。体外校正氯化物外排发生在高达25%的人类细胞中。深度测序显示部分同源位点的脱靶效应可以忽略不计。在CF小鼠鼻内应用纳米颗粒会产生鼻上皮电位差异的变化，与校正后的CFTR一致，在肺组织中也检测到基因校正。这项工作代表了利用纳米颗粒系统利用寡核苷酸进行体内基因组工程，实现临床相关水平的基因编辑而不产生脱靶效应。
Cystic fibrosis (CF) is a lethal genetic disorder most commonly caused by the F508del mutation in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. It is not readily amenable to gene therapy because of its systemic nature and challenges including in vivo gene delivery and transient gene expression. Here, we use triplex-forming PNA molecules and donor DNA in biodegradable polymer nanoparticles to correct F508del. We confirm modification with sequencing and a functional chloride efflux assay. In vitro correction of chloride efflux occurs in up to 25% of human cells. Deep sequencing reveals negligible off-target effects in partially homologous sites. Intranasal application of nanoparticles in CF mice produces changes in nasal epithelium potential differences consistent with corrected CFTR, with gene correction also detected in lung tissue. This work represents facile genome engineering in vivo with oligonucleotides using a nanoparticle system to achieve clinically relevant levels of gene editing without off-target effects.
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影响因子：
10
作者：
Fields RJ;Quijano E;McNeer NA;Caputo C;Bahal R;Anandalingam K;Egan ME;Glazer PM;Saltzman WM
通讯作者：
Saltzman WM
影响因子：
56.9
作者：
Egan, ME;Pearson, M;Caplan, MJ
通讯作者：
Caplan, MJ
影响因子：
14.8
作者：
Hutt DM;Herman D;Rodrigues AP;Noel S;Pilewski JM;Matteson J;Hoch B;Kellner W;Kelly JW;Schmidt A;Thomas PJ;Matsumura Y;Skach WR;Gentzsch M;Riordan JR;Sorscher EJ;Okiyoneda T;Yates JR 3rd;Lukacs GL;Frizzell RA;Manning G;Gottesfeld JM;Balch WE
通讯作者：
Balch WE
影响因子：
4.7
作者：
Akinc, A;Anderson, DG;Langer, R
通讯作者：
Langer, R
影响因子：
5.1
作者：
Bruscia, E;Sangiuolo, F;Gruenert, DC
通讯作者：
Gruenert, DC