Basic Information
Description
The exogenous promoter and chimeric MERTK coding sequence containing the human signal peptide, extracellular domain, transmembrane domain and mouse cytoplasmic domain was inserted to replace part of murine exon 1.
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Application
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B-hMERTK MC38 cells have the capability to establish tumors in vivo, which can be used for efficacy studies.
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Targeting Strategy
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The exogenous promoter and chimeric MERTK coding sequence containing the human signal peptide, extracellular domain, transmembrane domain and mouse cytoplasmic domain was inserted to replace part of murine exon 1. The insertion disrupts the endogenous murine Mertk gene, resulting in a non-functional transcript.
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Protein Expression Analysis
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Pre-inoculation
MERTK expression analysis in B-hMERTK MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hMERTK MC38 cell cultures were stained with species-specific anti-MERTK antibodies. Human MERTK was detected on the surface of B-hMERTK MC38 cells but not wild-type MC38 cells. The 1-G01 clone of B-hMERTK MC38 cells was used for in vivo experiments.
Post-inoculation
B-hMERTK MC38 cells were subcutaneously implanted into C57BL/6 mice (n=5). Tumor cells were harvested on day 34 post inoculation and assessed for species-specific MERTK expression by flow cytometry. As shown, human MERTK was highly expressed on the surface of humanized B-hMERTK MC38 tumor cells, indicating B-hMERTK MC38 cells can be used for in vivo efficacy studies of novel MERTK therapeutics.
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Tumor Growth Curve
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Tumor Growth Curve & Body Weight Changes in C57BL/6N Mice
Subcutaneous homograft tumor growth of B-hMERTK MC38 cells. B-hMERTK MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into C57BL/6N mice (female, 7-week-old, n=5). (A) Average tumor volume ± SEM and (B) body weight (mean ± SEM) were measured twice a week. Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hMERTK MC38 cells were able to establish tumors in vivo, which can be used for efficacy studies.
