The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesi...The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesis and transgenic methods to study gene function and improve crop traits. CRISPR/Cas facilitates targeted gene editing through RNA-guided DNA cleavage followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. Here we describe a detailed procedure for our previously developed and highly efficient CRISPR/Cas9 method that allows the generation of heritable-targeted gene mutations andcorrections in Arabidopsis. This protocol describes the strategies and steps for the selection of targets, design of single-guide RNA(sg RNA), vector construction and analysis of transgenic lines. We also offer a method to target two loci simultaneously using vectors containing two different sg RNAs. The principles described in this protocol can be applied to other plant species to generate stably inherited DNA modifications.展开更多
It is shown that the nonautonomous discrete Toda equation and its Backlund transformation can be derived from the reduction of the hierarchy of the discrete KP equation and the discrete two-dimensional Toda equation. ...It is shown that the nonautonomous discrete Toda equation and its Backlund transformation can be derived from the reduction of the hierarchy of the discrete KP equation and the discrete two-dimensional Toda equation. Some explicit examples of the determinant solutions of the nonautonomous discrete Toda equation including the Askey-Wilson polynomial are presented. Finally we discuss the relationship between the nonautonomous discrete Toda system and the nonautonomous discrete Lotka- Volterra equation.展开更多
基金supported by the Chinese Academy of Sciences and China Scholarship Council(201206050103)
文摘The newly developed CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas(CRISPR-associated) system has emerged as an efficient tool for genome-editing, providing an alternative to classical mutagenesis and transgenic methods to study gene function and improve crop traits. CRISPR/Cas facilitates targeted gene editing through RNA-guided DNA cleavage followed by cellular DNA repair mechanisms that introduce sequence changes at the site of cleavage. Here we describe a detailed procedure for our previously developed and highly efficient CRISPR/Cas9 method that allows the generation of heritable-targeted gene mutations andcorrections in Arabidopsis. This protocol describes the strategies and steps for the selection of targets, design of single-guide RNA(sg RNA), vector construction and analysis of transgenic lines. We also offer a method to target two loci simultaneously using vectors containing two different sg RNAs. The principles described in this protocol can be applied to other plant species to generate stably inherited DNA modifications.
基金supported in part by Grant-in-Aid for Scientific Research No. 18540214 from the Ministry of Education, Culture, Sports, Science, and Technology, Japan
文摘It is shown that the nonautonomous discrete Toda equation and its Backlund transformation can be derived from the reduction of the hierarchy of the discrete KP equation and the discrete two-dimensional Toda equation. Some explicit examples of the determinant solutions of the nonautonomous discrete Toda equation including the Askey-Wilson polynomial are presented. Finally we discuss the relationship between the nonautonomous discrete Toda system and the nonautonomous discrete Lotka- Volterra equation.
