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The mixture of hA3A-eBE3-Y130F mRNA (100 ng/L) and Trim37- sgRNA-stop (100 ng/L) was diluted in RNase-free water, centrifuged at 13,400 g for 10 min at 4 C, and injected into the cytoplasm of zygotes ( mTmG Blimp1-T2A-Cre or B6D2F1 Nanog-EGFP ) using a micromanipulator (Olympus, Tokyo, Japan) and a FemtoJet microinjector (Eppendorf)
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Continue normal diet emphasizing protein and nutrient density
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