Gene Function Studies

Use conventional knockout when gene deletion does not cause embryonic lethality and you want to study global gene function throughout development and adult life. Use conditional knockout when genes are essential for development (would cause lethality), when you need tissue specific deletion, or when avoiding developmental compensation is important.

Use conditional knockout with tissue specific Cre drivers (e.g., LysM-Cre for macrophages, Albumin-Cre for hepatocytes, Nestin-Cre for neural cells). This enables study of gene function in specific cell populations without systemic effects. Validate Cre driver specificity and recombination efficiency before experimental studies.

Yes. Generate double-knockout mice (both genes deleted) to study gene interactions, epistasis, and genetic hierarchies. Breeding requires careful planning to combine alleles. Conditional approaches enable cell type specific study of multiple genes by using different Cre drivers or combining Cre with other recombinase systems (FLP, Dre).

Essential controls include wild-type littermates (genetic background control), heterozygous carriers (dose-response), Cre-positive flox-negative controls (to detect Cre toxicity), and vehicle-treated controls (for inducible systems). Reporter crosses validate recombination efficiency. Multiple controls ensure phenotypes are due to gene deletion, not experimental artifacts.