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The I123H/D195K/D208R/V232A mutant of Cas12f from Acidibacillus sulfuroxidans (described as Cas12f in this paper) 52 , Halo7-tag, and FRB were inserted after the CMV promotor of the vector, and followed by U6 promoter for expressing sgRNA for the mouse transthyretin (TTR) gene (as guide RNA) 52 , resulting in pKLV2-AsCas12fHKRA-Halo7-GSGS-FRB-guideRNA

The insertion of amino acids in the GFP backbone may lead to changes in the spectroscopic properties of GFP but still yield functional fluorescent proteins (Topell et al., 2 and Table 1)