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Gene Synthesis> | Codon-optimized anti-human CD47 scFv (B6H12) DNA was synthesized by GenScript (Piscataway, NJ) . and differentiated into macrophages in the presence of 100 ng/mL M-CSF7 (GenScript, NJ, USA). A single guide RNA (sgRNA) targeting exon 1 of the human CD47 gene (guide sequence: GGAGATGTGGCCCCTGGTAG) was synthesized by Genscript Biotech (Piscataway, NJ, USA). A total of 30 pmol of the synthesized sgRNA was incubated with 15 pmol of GenCRISPR™ Cas9 v1.2 nuclease (Genscript Biotech) in 25 µl of serum-free Opti-MEM medium (ThermoFisher, Waltham, MA, USA) for 10 minutes at room temperature to form a ribonucleoprotein (RNP) complex. | Get A Quote |
CD47 is overexpressed on the surface of many types of cancer cells, including T-cell acute lymphoblastic leukemia (T-ALL) cells. In this study, we have developed a diphtheria toxin-based bivalent anti-human CD47 immunotoxin (bi-CD47-IT) for the targeted therapy of CD47+ cancers using a unique diphtheria toxin-resistant yeast Pichia pastoris expression system. Bi-CD47-IT demonstrated compelling in vivo efficacy in multiple T-ALL cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) mouse models. Bi-CD47-IT significantly prolonged the median survival of the tumor-bearing mice and highly effectively depleted the T-ALL blast cells in the peripheral blood, spleen, liver, bone marrow, brain, and spina... More