Method for knocking off myo7ab gene of zebra fish
A zebrafish and gene technology, applied in the field of gene knockout, can solve the problems of high cost, many steps, time-consuming, reagent and sequencing costs, etc.
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[0043] 1) Design CRISPR / Cas9 gene knockout target sites and detection primers
[0044] Query the genomic DNA sequence of the zebrafish myo7ab gene on the National Center for Biotechnology Information (NCBI), analyze its functional domain on the website SMART (http: / / smart.embl-heidelberg.de / ), and knock out it according to CRISPR / Cas According to the principle, the target site of the myo7ab gene was designed on the website The ZiFiT Targeter (http: / / zifit.partners.org / ZiFiT / ). The selection of targets must follow this standard: 5'-GG-(N)18-NGG-3'. The GG dinucleotide at the 5' end is part of the T7 promoter, and this restriction is not required when designing the target site, but it must be ensured that the 3' end of the target site is NGG. The selected position of the target must be within the structural domain of the gene to ensure that the insertion or deletion of bases at the target site can affect the entire structural domain of the myo7ab gene, thereby changing the expr...
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