Method for creating fragrant rice by knocking out BADH2 (betaine aldehyde dehydrogenase) gene by utilizing CRISPR/Cas9 system
A systematic knockout and gene technology, applied in genetic engineering, plant gene improvement, chemical instruments and methods, etc., can solve the problem that gene functions cannot be completely removed, and achieve the effect of shortening the breeding period and simple operation of the method
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[0032] Specific embodiments of the present invention will be described in detail below.
[0033] A method for creating fragrant rice by knocking out the BADH2 gene using the CRISPR / Cas9 system, comprising the steps of:
[0034] (1) Design the sgRNA sequence based on the BADH2 gene
[0035] Use the online public software CRISPR-P (http: / / cbi.hzau.edu.cn / crispr / ) to design the sgRNA guide sequence, select the guide sequence with a higher score and located at the appropriate position of the BADH2 gene sequence, the one selected in this example The sgRNA guide sequence is the sequence shown in SEQ ID No.2 and SEQ ID No.3.
[0036] (2) Design and synthesize sgRNA primers
[0037] According to the original sgRNA sequence and vector construction requirements designed by the online software, for the sgRNA sequence shown in SEQ ID No.2, the designed forward and reverse primers are sgU6-F
[0038] (cttgACTAGAGACGCTTGATTGT) and sgU6-R
[0039] (aaacACAATCAAGCGTCTCTAGT); for the sgRNA...
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