Leiden University Medical Center, Department of Molecular Cell Biology, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.
Leiden University Medical Center, Department of Molecular Cell Biology, Einthovenweg 20, 2333 ZC, Leiden, The Netherlands.
Trends Biotechnol. 2015 May;33(5):280-91. doi: 10.1016/j.tibtech.2015.02.011. Epub 2015 Mar 26.
Genome editing (GE) entails the modification of specific genomic sequences in living cells for the purpose of determining, changing, or expanding their function(s). Typically, GE occurs after delivering sequence-specific designer nucleases (e.g., ZFNs, TALENs, and CRISPR/Cas9) and donor DNA constructs into target cells. These designer nucleases can generate gene knockouts or gene knock-ins when applied alone or in combination with donor DNA templates, respectively. We review progress in this field, with an emphasis on designer nuclease and donor template delivery into mammalian target cell populations. We also discuss the impact that incremental improvements to these tools are having on the specificity and fidelity attainable with state-of-the-art DNA-editing procedures. Finally, we identify areas that warrant further investigation.
基因组编辑（GE）涉及对活细胞中的特定基因组序列进行修饰，目的是确定、改变或扩展其功能。通常，GE 是在将序列特异性设计核酸酶（例如 ZFNs、TALENs 和 CRISPR/Cas9）和供体 DNA 构建体递送至靶细胞之后进行的。这些设计核酸酶在单独或与供体 DNA 模板组合使用时可以分别产生基因敲除或基因敲入。我们回顾了该领域的进展，重点介绍了设计核酸酶和供体模板递送至哺乳动物靶细胞群体的情况。我们还讨论了这些工具的逐步改进对最新 DNA 编辑程序的特异性和保真度的影响。最后，我们确定了需要进一步研究的领域。
