Efficient generation of endogenous protein reporters for mouse development

Fluorescent proteins and epitope tags can reveal protein localization in cells animals, yet the large size of many hinders efficient genome targeting. Accordingly, studies have relied on characterizing overexpressed proteins, which might not recapitulate endogenous activities. Here, we present two strategies for higher throughput production reporters mice, focusing blastocyst model development. Our first strategy makes use a split fluorescent mNeonGreen2 (mNG2). Knock-in small portion mNG2 gene, frame with gene coding regions interest was highly embryos, potentially obviating need to establish mouse lines. When complemented by larger fluorescence reconstituted faithfully reported living embryos. second achieves in-frame knock-in relatively tag, provides high efficiency sensitivity reporting. Together, these approaches provide complementary advantages enable broad downstream applications.