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Overview
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AapCas12b (previously known as AapC2c1) is a DNA endonuclease mediated by tracrRNA and crRNA (or sgRNA), which can specifically cleave the dsDNA target in a PAM (TTN)-dependent manner, generating DSBs, and cleave ssDNA target in a PAM-independent manner. In addition, both dsDNA and ssDNA targets can trigger the trans-cleavage activity of AapCasI2b, that is, when Cas12b binds sgRNA and target DNA to form a ternary complex, the trans-cleavage activity of Cas12b will be released to non-specifically cleave ssDNA sequences in the reaction system. AapCas12b is derived from Alicyclobacillus acidophilus. The optimal reaction temperature is 60°C for AapCas12b. Therefore, compared to AacCas12b, AapCas12b is more suitable for “One-Step” CRISPR-Dx system with the integration of LAMP isothermal amplification (for details, please refer to PMID: 32937062).
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Overview