Mutants in genes with no visible phenotype required PCR screening for their identification; therefore, high mutagenesis rates are important, which might be difficult to achieve at all positions in the fly genome (Kondo and Ueda 2013; Yu et al. 2013, 2014; Gokcezade et al. 2014; Port et al. 2014).
The presence of such inactive protease variants in plant genomes suggests that they might have acquired roles other than proteolysis, resulting in altered evolutionary pressure on the protease domain and the potential for higher mutagenesis rates.
Since the apicoplast genome is contained on 35 kb of circular DNA [ 65] this indicates a 100-fold higher spontaneous mutagenesis rate in the plastid's genome than in the nuclear genome.
Finally, we generated null alleles of HP1a using optimized parameters through homology-directed repair and achieved an overall mutagenesis rate significantly higher than previously reported.
We found that <3 to 4 days of administration of the virus, the mutagenesis rate of Pcsk9 in the liver was as high as >50%.
The calculated mutagenesis rates are 91- and 2000-fold higher than the spontaneous mutation rate of 6.0 × 10−9 mutations/effective site calculated for Arabidopsis.
Mutagenesis rates were calculated using the Drake Formula [65].
act-cas9, vasa-cas9 BL51323, and nos-cas9 TH00787.N (which we refer to as TH_attP2) gave rise to similar, high rates of mutagenesis.
The aforementioned results, together with previous literature (Kondo and Ueda 2013; Port et al. 2014), provide evidence that transgenic supply of both Cas9 and gRNA can lead to very high rates of mutagenesis.
In addition, Ubc12 knockout enhanced PCNA-polη interaction, but NEDD8 expression or NEDP1 deletion reduced the recruitment of polη (Fig. 6), indicating that enhanced NEDDylation reduces excessive polη staying on the replication fork to avoid high rate of mutagenesis.
