J Biotechnol:南京农业大学喻德跃课题组用基因组编辑技术对大豆高

基因打靶(GT)对于推进基础植物学研究和作物改良，具有重要的意义。TALENs和CRISPR/Cas9系统已被开发用于真核生物的基因组编辑，包括植物。在这项研究中，研究人员对两个大豆基因组编辑靶标——GmPDS11和GmPDS18，进行了这两种技术的比较分析。研究人员发现，对于大豆毛状根中的基因打靶，利用TALENs技术的单个打靶效率范围在17.5%至21.1%，而使用AtU6-26promoter的CRISPR/Cas9效率范围在11.7%到18.1%，使用GmU6-16g-1promoter的CRISPR/Cas9效率范围在43.4%至48.1%，从而表明，相比较他技术而言，使用gmu6-16g-1promoter的CRISPR/Cas9，可能是一种更有效的工具。

同样的，研究人员对着三种技术进行的双突变GT效率实验显示，TALENs的打靶效率为6.25%，使用atu6-26promoter 的CRISPR/Cas9打靶效率为12.5%，使用gmu6-16g-1promoter的CRISPR/Cas9打靶效率为43.4%至48.1%，从而表明，CRISPR/Cas9仍是同时编辑多个同源等位基因(homoeoallele)的一个更好选择。而且，研究人员在子叶节点中观察到了大豆转化引起的白化和矮芽(PDS基因敲除)。

总而言之这些研究结果表明，TALENs和CRISPR/Cas9系统是大豆基因组编辑的一种强大工具。而且，GmU6-16g-1是大豆sgRNA表达的一种合适的U6promoter。

原文链接：

Efficient targeted mutaGENEsis in soybean by TALENs and CRISPR/Cas9

原文摘要：

Gene targeting (GT) is of great significance for advancing basic plant research and crop improvement. Both TALENs (transcription activator-like effectors nucleases) and CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated 9) systems have been developed for genome editing in eukaryotes, including crop plants. In this work, we present the comparative analysis of these two technologies for two soybean genome editing targets, GmPDS11 and GmPDS18. We found GT in soybean hairy roots with a single targeting efficiency range of 17.5% to 21.1% by TALENs, 11.7% to 18.1% by CRISPR/Cas9 using the AtU6-26promoter, and 43.4% to 48.1% by CRISPR/Cas9 using the GmU6-16g-1promoter, suggesting that the CRISPR/Cas9 using the GmU6-16g-1promoter is probably a much more efficient tool compared to the other technologies. Similarly, our double mutation GT efficiency experiment with these three technologies displayed a targeting efficiency of 6.25% by TALENs, 12.5% by CRISPR/Cas9 using the AtU6-26promoter, and 43.4% to 48.1% by CRISPR/Cas9 using the GmU6-16g-1promoter, suggesting that CRISPR/Cas9 is still a better choice for simultaneous editing of multiple homoeoalleles. Furthermore, we observed albino and dwarf buds (PDS knock-out) by soybean transformation in cotyledon nodes. Our results demonstrated that both TALENs and CRISPR/Cas9 systems are powerful tools for soybean genome editing.

doi: 10.1016/j.jbiotec.2015.11.005.

作者：喻德跃