CRISPR–Cas9 methods have been applied to generate random insertions and deletions, large deletions, targeted insertions or replacements of short sequences, and precise base changes in plants^{1,2,3,4,5,6,7}. However, versatile methods for targeted insertion or replacement of long sequences and genes, which are needed for functional genomics studies and trait improvement in crops, are few and largely depend on the use of selection markers^8,9,10,11. Building on methods developed in mammalian cells¹², we used chemically modified donor DNA and CRISPR–Cas9 to insert sequences of up to 2,049 base pairs (bp), including enhancers and promoters, into the rice genome at an efficiency of 25%. We also report a method for gene replacement that relies on homology-directed repair, chemically modified donor DNA and the presence of tandem repeats at target sites, achieving replacement with up to 130-bp sequences at 6.1% efficiency.

CRISPR–Cas9方法已应用于在植物^{1,2,3,4,5,6,7中}产生随机插入和缺失，大缺失，短序列的目标插入或替换以及精确的碱基改变。但是，针对功能性基因组学研究和农作物性状改良所需的长序列和基因定向插入或替换的通用方法很少，并且在很大程度上取决于选择标记^8,9,10,11的使用。建立在哺乳动物细胞中的方法上¹²，我们使用化学修饰的供体DNA和CRISPR–Cas9将高达2049个碱基对（bp）的序列（包括增强子和启动子）插入水稻基因组，效率为25％。我们还报告了一种基因置换的方法，该方法依赖于同源性指导的修复，化学修饰的供体DNA和在目标位点的串联重复序列的存在，以6.1％的效率实现了高达130 bp的序列置换。