CRISPR-Cas9 system for preventing and/or treating HIV, as well as preparation method and application thereof

[0113] (1) Acquisition of sgRNA gene

[0114] The sgRNAs primer sequences were directly ordered from Invitrogen, and the relevant sequences are shown in Table 1.

[0115] Table 1: sgRNA primer sequences

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[0121] (2) Construct sgRNA into expression vector

[0122] a): PCR amplification of sgRNA

[0123] Reaction system: sterile ddH 2 O: 37.5 μL, 10×PCR Buffer (containing MgCl 2 ): 5 μL, 2.5 mM dNTP: 4 μL, corresponding upstream and downstream primers 1 μL each (primer concentration 50 pmol / μL), template DNA (50 ng / μL): 1 μL, PyrobestTM DNA Polymerase: 0.5 μL. After gentle shaking and mixing, PCR amplification was carried out according to the conditions shown in Table 2.

[0124] Table 2: PCR Amplification Conditions

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[0127] b) Vector construction and identification

[0128] Use the PCR recovery kit produced by Quigen to recover the PCR amplification product according to the conventional