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Future research is warranted to explore this dosing strategy in larger patient populations

One week of geneticin selection post-nucleofection resulted in stable expression of both sensors (Fig 4B)

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sgRNA constructs and gene expressing codon optimized (OPT) plasmid constructs sgRNA control and constructs against CBS and L2HGDH were designed targeting GTATTACTGATATTGGTGGG (sgControl) (#230080, Addgene), GGAGCAGACAACCTACGAGG (sg CBS -1) (#230081, Addgene), TGGCAGTGCTGGCAGCACGG (sg CBS -2) (#230082, Addgene), GATATAGTCATCGTTGGTGG (sg L2HGDH -1) (#230083, Addgene), and CACCAGACTGGACATAACAG (sg L2HGDH -2) (#230084, Addgene) and constructed in lentiCRISPR v2-Blast (#83480, Addgene) for Blasticidin selection at a final concentration of 5 g/ml