Construction method of FNDC5 gene knock-out rat model, and application
A technology of gene knockout and rat model, applied in the construction of FNDC5 gene knockout rat model, field of gene knockout
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[0029] 1. Design of gene knockout model rats
[0030] like figure 1 As shown, the FNDC5 gene is on the positive strand of chromosome 5, with a total length of about 7kb. Gene ID: 260327. like figure 2 As shown, the gene has 1 transcript, and this design refers to transcript 201. The FNDC5 gene sequence is shown in SEQ ID NO.8.
[0031] Analysis of the FNDC5 gene structure, selection to delete Exon2-6. Two sgRNAs were designed on Intron1 and 3'UTR respectively, resulting in about 4kb base deletion, so as to achieve the purpose of FNDC5 knockout. Model rats were prepared using the CRISPR / Cas9 system. Knockout strategies such as image 3 shown.
[0032] 2. Preparation of gene knockout rat model
[0033] 2.1 Cas9 / sgRNA design and activity detection
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