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WT and ACLY KO cells were then transduced with a LentiCRISPR v2 vector with no guide RNA or containing a guide RNA targeting the first exon of ACSS2 or near the active site of CrAT: ACSS2 KO, DKO1, DKO2 (and KPACs): mACSS2sg2 - CGAGCTGCACCGGCGTTCTG DKO3: mACSS2sg6 - CTGCACCGGCGTTCTGTGG ACLY KO CrAT KO1, ACLY KO CrAT KO2: msgCRAT1F - CACCGTCCACAAGTGCAACTATGGG Following transduction, cells were treated with puromycin until the entirety of an un-transduced cell population died

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Result: Smoother motion, less stiffness, and lasting joint protection

Chemical & Physical Properties Toxicological Information CHEMICAL IDENTIFICATION RTECS NUMBER : MC0556000 CHEMICAL NAME : Glycine, N-(N-L-gamma-glutamyl-L-cysteinyl)- CAS REGISTRY NUMBER : 70-18-8 LAST UPDATED : 199710 DATA ITEMS CITED : 20 MOLECULAR FORMULA : C10-H17-N3-O6-S MOLECULAR WEIGHT : 307.36 HEALTH HAZARD DATA ACUTE TOXICITY DATA TYPE OF TEST : LD50 - Lethal dose, 50 percent kill ROUTE OF EXPOSURE : Oral SPECIES OBSERVED : Rodent - mouse DOSE/DURATION : 5 gm/kg TOXIC EFFECTS : Details of toxic effects not reported other than lethal dose value TYPE OF TEST : LD50 - Lethal dose, 50 percent kill ROUTE OF EXPOSURE : Intraperitoneal SPECIES OBSERVED : Rodent - mouse DOSE/DURATION : 4020 mg/kg TOXIC EFFECTS : Details of toxic effects not reported other than lethal dose value TYPE OF TEST : LD50 - Lethal dose, 50 percent kill ROUTE OF EXPOSURE : Subcutaneous SPECIES OBSERVED : Rodent - mouse DOSE/DURATION : 5 gm/kg TOXIC EFFECTS : Details of toxic effects not reported other than lethal dose value TYPE OF TEST : LD50 - Lethal dose, 50 percent kill ROUTE OF EXPOSURE : Intravenous SPECIES OBSERVED : Rodent - mouse DOSE/DURATION : 2238 mg/kg TOXIC EFFECTS : Details of toxic effects not reported other than lethal dose value TYPE OF TEST : LD50 - Lethal dose, 50 percent kill ROUTE OF EXPOSURE : Intramuscular SPECIES OBSERVED : Rodent - mouse DOSE/DURATION : 4 gm/kg TOXIC EFFECTS : Details of toxic effects not reported other than lethal dose value TYPE OF TEST : TDLo - Lowest published toxic dose ROUTE OF EXPOSURE : Oral DOSE : 1250 mg/kg SEX/DURATION : female 1-22 day(s) after conception TOXIC EFFECTS : Reproductive - Fertility - post-implantation mortality (e.g