A simple,fast and efficient method of constructing dual-target CRISPR/Cas9 vector
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Abstract:
A simple,fast and efficient method of constructing dual-target CRISPR/Cas9 vector was established based on the Golden Gate ligation. The two designed small target DNA fragments and one exogenous fragment were constructed together into the CRISPR/Cas9 vector with T4 DNA ligase. 20 dual-target CRISPR/Cas9 vectors against 10 reported rice genes were constructed with this method,and the positive rate was 100% with no mutation in sequences. This vector system provides an easy,rapid and efficient method for constructing dual-target CRISPR/Cas9 vector for rice. It also provides a reference for the construction of dual-target CRISPR/Cas9 vectors in other plants.
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张扬,黄维峰,周菲,林拥军. A simple, fast and efficient method of constructing dual-target CRISPR/Cas9 vector [J]. Jorunal of Huazhong Agricultural University,2020,39(3):9-18.