The following information accompanied stocks donated to the Bloomington Drosophila Stock Center by Joshua Mast and Nina DiPrimio, Perlara PBC.
CG14291L89P - CRISPR/Cas9-mediated homologous dependent repair was used to make amino acid change L89P. CG14291 is the fly ortholog of human N-sulfoglucosamine sulfohydrolase (SGSH), a sulfatase. The L89P change was put in along with a 3xP3-dsRed marker flanked by PBac ends that was then removed via PBac transposition. Transposition left behind a TTAA which changes CTGAAC to CTTAAC but doesn't change the amino acid sequence. L89P was likely chosen based on the L91P change found in a human N-acetylgalactosamine-6-sulfatase (GALNS) variant associated with Mucopolysaccharidosis type 4A. GALNS is also a sulfatase and L91P is thought to knock out enzymatic activity.
CG14291S301P - CRISPR/Cas9-mediated homologous dependent repair was used to make amino acid change S301P designed to mimic the S298P change found in a human N-sulfoglucosamine sulfohydrolase (SGSH) variant associated with Mucopolysaccharidosis type IIIA (Sanfilippo syndrome A). The S301P change was put in along with a 3xP3-dsRed marker flanked by PBac ends that was then removed via PBac transposition. Transposition left behind a TTAA but doesn't change the amino acid sequence.
CG14291S64W - CRISPR/Cas9-mediated homologous dependent repair was used to make amino acid change S64W designed to mimic the S66W change found in a human N-sulfoglucosamine sulfohydrolase (SGSH) variant associated with Mucopolysaccharidosis type IIIA (Sanfilippo syndrome A). The S64W change was put in along with a 3xP3-dsRed marker flanked by PBac ends that was then removed via PBac transposition. Transposition left behind a TTAA but doesn't change the amino acid sequence.