Transformation: OsRT0004

Transformation Information

Transformation Name: OsRT0004
Description: transformation OsRT0004 to Nipponbare or Kasalath
Genetic background: Nipponbare or Kasalath
Project Name: The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation
Description: 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.

Construct for Transformation (OsRT0004)

Plants for Transformation (OsRT0004)

Note: The tree nodes can be clicked to show the generations of plants. T0 represents the 1st generation (Progenitor of T1), T1 represents the 2nd generation (Progeny of T0), and so on. Clicking the plant name will be directed to the plant information page.

This database is supported by NSF (IOS-1546625) and hosted by BTI.