Correction of chromosomal point mutations in human cells with bifunctional oligonucleotides
Correction of chromosomal point mutations in human cells with bifunctional oligonucleotides
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DOI：
10.1038/13684
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发表时间：
1999-10-01
影响因子：
46.9
通讯作者：
Khorlin, A
中科院分区：
文献类型：
--
作者：
Culver, KW;Hsieh, WT;Khorlin, A
通过将两个不同的结合结构域整合到单链寡核苷酸中，设计了一种用于染色体突变纠正的序列特异性基因组传递系统。一个修复结构域（RD）包含目标区域的原生序列，第三个链形成结构域（TFD）被设计成Hoogsteen相互作用形成三倍体。设计的前提是RD会迅速形成异源双工，与TFD协同锚定，脱氧寡核苷酸被设计成在人类腺苷脱氨酶（ADA）和p53基因附近形成三联体，邻近已知的点突变，转染ADA缺陷的人类淋巴细胞纠正了1-2%的细胞突变序列。RD和TFD都没有单独纠正突变，转染p53突变的人胶质母细胞瘤细胞纠正了突变，并诱导7.5%的细胞凋亡。
A sequence-specific genomic delivery system for the correction of chromosomal mutations was designed by incorporating two different binding domains into a single-stranded oligonucleotide. A repair domain (RD) contained the native sequence of the target region, A third strand-forming domain (TFD) was designed to form a tripler by Hoogsteen interactions. The design was based upon the premise that the RD will rapidly form a heteroduplex that is anchored synergistically by the TFD, Deoxyoligonucleotides were designed to form triplexes in the human adenosine deaminase (ADA) and p53 genes adjacent to known point mutations, Transfection of ADA-deficient human lymphocytes corrected the mutant sequence in 1-2% of cells. Neither the RD or TFD individually corrected the mutation, Transfection of p53 mutant human glioblastoma cells corrected the mutation and induced apoptosis in 7.5% of cells.