Plant Crispr Database

Project Information

The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation

The CRISPR/Cas9 system has been demonstrated to efficiently induce targeted gene editing in a variety of organisms including plants. Recent work showed that CRISPR/Cas9-induced gene mutations in Arabidopsis were mostly somatic mutations in the early generation, although some mutations could be stably inherited in later generations. However, it remains unclear whether this system will work similarly in crops such as rice. In this study, we tested in two rice subspecies 11 target genes for their amenability to CRISPR/Cas9-induced editing and determined the patterns, specificity and heritability of the gene modifications. Analysis of the genotypes and frequency of edited genes in the first generation of transformed plants (T0) showed that the CRISPR/Cas9 system was highly efficient in rice, with target genes edited in nearly half of the transformed embryogenic cells before their first cell division. Homozygotes of edited target genes were readily found in T0 plants. The gene mutations were passed to the next generation (T1) following classic Mendelian law, without any detectable new mutation or reversion. Even with extensive searches including whole genome resequencing, we could not find any evidence of large-scale off-targeting in rice for any of the many targets tested in this study. By specifically sequencing the putative off-target sites of a large number of T0 plants, low-frequency mutations were found in only one off-target site where the sequence had 1-bp difference from the intended target. Overall, the data in this study point to the CRISPR/Cas9 system being a powerful tool in crop genome engineering.

Lab: Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences, Shanghai, China and Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, USA
Contact: Nanfei Xu (nfxu@sibs.ac.cn) and Jian-Kang Zhu (jkzhu@purdue.edu)

List of Transformation

Transformation	Description	Genetic Background	Construct	Publication
OsRT0001	transformation OsRT0001 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-1	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0002	transformation OsRT0002 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-2	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0003	transformation OsRT0003 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-3	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0004	transformation OsRT0004 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-4	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0005	transformation OsRT0005 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-5	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0006	transformation OsRT0006 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-6	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0007	transformation OsRT0007 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-7	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0008	transformation OsRT0008 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-8	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.
OsRT0009	transformation OsRT0009 to Nipponbare or Kasalath	Nipponbare or Kasalath	OsTG0001-9	Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y.,Yang, L., Zhang, H., Xu, N., et al. (2014). The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12:797每807.