engleski

Generation of new mouse models for DNA damage response

Programmable nucleases are a method of choice for creation of mouse models where precision is required, for example in creating fluorescent fusion proteins under the control of the endogenous promoter to study protein function under physiological conditions in vivo. Most of the studies using fluorescently tagged proteins use expression vectors, and therefore create an unnatural overabundance of the protein, which may affect its activity. Here we present insertion of citrine gene after the ATG site of DNA damage response genes Fancd2 and 53BP1 using TALENs and CRISPRs. The correct integration was verified by southern blot, and functionality was demonstrated by induction of DNA damage in mouse embryonic fibroblasts.

DNA damage; gene editing nucleases; mouse model

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