Figure 1.
Overview of the retroviral mutagenesis pipeline. The pseudotyped murine leukemia virus (A) is injected into 1000–2000 cell stage blastula embryos (B). The infection rate is determined by quantitative PCR (qPCR), and founder fish with high infection rates are raised to adults. The founders are crossed to wild-type (T/AB) fish, and F1 male fish are used for sperm cryopreservation and fin biopsies. Integrations are amplified and mapped from gDNA isolated from the fin biopsies. Mapped integrations are assigned to the corresponding sperm samples, and desired mutations are recovered by in vitro fertilization.
