Comparison of CRISPR/Cas9 and TALENs on editing an integrated EGFP gene in the genome of HEK293FT cells.
Comparison of CRISPR/Cas9 and TALENs on editing an integrated EGFP gene in the genome of HEK293FT cells.
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DOI：
10.1186/s40064-016-2536-3
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
Lillico SG
中科院分区：
文献类型：
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作者：
He Z;Proudfoot C;Whitelaw CB;Lillico SG
诸如CRISPR/CAS9和TALENS等基因组编辑程序处于哺乳动物基因组靶向修饰方法研究的前沿。到目前为止，很少有人对CRISPR/CAS9和TALENS的基因组编辑特性的差异进行比较研究。虽然CRISPR/CAS9系统已经取代TALEN成为从事这一领域工作的大多数研究小组的首选工具，但我们假设，TALEN的应用将会有特定的好处。在这里，我们比较了CRISPR/CAS9和TALEN作为工具，在HEK293FT细胞基因组中引入整合的EGFP基因的位点特异性编辑事件。GUIDE RNAs和TALEN对是针对EGFP基因内的两个基因座设计的。我们发现，与两个TALEN对相比，配对的Cas9核酸酶更有效和更精确地诱导靶向基因组缺失。然而，当同时提供跨越EGFP内两个DNA双链断裂(DSB)的质粒模板时，TALENS比CRISPR/Cas9更有效地刺激同源定向修复(HDR)，并导致更少的靶向基因组缺失。我们的数据表明，基因组编辑工具的选择应该取决于期望的基因组编辑结果。这样一种理性的方法可能会使从事各种领域工作的团体的研究成果受益，这些领域包括细胞系修改、疾病研究的动物模型或基因治疗策略。本文的在线版本(doi：10.1186/s40064-0162536-3)包含补充材料，可供授权用户使用。
Genome editors such as CRISPR/Cas9 and TALENs are at the forefront of research into methodologies for targeted modification of the mammalian genome. To date few comparative studies have been carried out to investigate the difference of genome editing characteristics between CRISPR/Cas9 and TALENs. While the CRISPR/Cas9 system has overtaken TALENs as the tool of choice for most research groups working in this field, we hypothesized that there could be certain applications whereby the application of TALENs would have specific benefits. Here we compare CRISPR/Cas9 and TALEN as tools for introducing site-specific editing events at an integrated EGFP gene in the genome of HEK293FT cells. Guide RNAs and TALEN pairs were designed to target two loci within the EGFP gene. We found that paired Cas9 nucleases induced targeted genomic deletion more efficiently and precisely than two TALEN pairs. However, when concurrently supplied with a plasmid template spanning the two DNA double-strand breaks (DSBs) within EGFP, TALENs stimulated homology directed repair (HDR) more efficiently than CRISPR/Cas9 and caused fewer targeted genomic deletions. Our data suggest that the choice of genome editing tool should be determined by the desired genome editing outcome. Such a rational approach is likely to benefit research outputs for groups working in fields as diverse as modification of cell lines, to animal models for disease studies, or gene therapy strategies. The online version of this article (doi:10.1186/s40064-016-2536-3) contains supplementary material, which is available to authorized users.