Fig. 1.

Isolation and characterization of TALEN-induced mutations in C. briggsae. (A) Many F₁ animals were heterozygous for two distinct mutations. Following injection of TALENs targeting the Cbr-him-8 gene, we isolated an F₁ animal with the Him phenotype (high incidence of male progeny). DNA from eight F₂ self-progeny was amplified by the polymerase chain reaction (PCR) and separated on a 10% acrylamide gel, revealing the presence of two different alleles. The sequence of each lesion is shown at the right. (B) TALENs can produce large deletion mutations in nematodes. DNA from potential Cbr-unc-51 mutants was amplified using four sets of primer pairs (a–d). The size of each expected fragment is indicated in (D). Mutant Cbr-unc-51(v215) lacked DNA in the region of primer pairs c and d. (C) The primer pair e could amplify a single band of approximately 3 kb from v215 DNA, indicating that most of this region was deleted. (D) Map of the region, showing the extent of the 9423 bp v215 deletion, determined by sequencing the fragment shown in (C). Positions on the map are related to the start of Cbr-unc-51.