BIOL 14a Lecture Notes - Lecture 28: Nucleic Acid Hybridization, Wild Type, Blastocyst
16 Nov 2016
School
Department
Course
Professor
Document Summary
Gene targeting: homologous vs. nonhomologous recombination both happen naturally. Doesn"t necessarily need to be targeted, so may not have to use embryonic stem cells (escs) Targeted injection: select es cells are added to an embryo to make a chimeric mouse. We are adding a gene we want to study or use in the animal (e. g. , the gfp mice) If non-targeted, we can modify a zygote directly instead of using escs. Dna is injected into pronucleus of recently fertilized egg. Surrogate mother mouse and her fully- transgenic offspring. No control over where transgenic dna is integrated into genome! Use a highly specific nuclease to cut dna in a specific, targeted gene damage that one gene. Specific to one gene (more specific than restriction enzyme) Rely on cells dna repair pathways to put the dna back together. Use a sequence-specific endonuclease to make targeted mutations in a genome. Special feature of cas9: sequence-specificity is determined primarily by:
