Effect of Sc contents on the mechanical properties and damping capacity of Al-Zn-Mg-Cu-Zr-Sc alloys with different grain structures.

Al-Zn-Mg-Cu-Zr-Sc alloys with different Sc contents were fabricated by casting, deformation, and T6 treatment. Deformation methods including rolling and friction-stir processing (FSP) were used to design their grain structure. A low additive amount (0.1) of Sc cannot refine the grains of the alloy with rolling and T6 treatment, and it instead coarsened the grains. The reason was the non-uniform distribution of nanosize Al3(Sc,Zr) phases that led to the occurrence of abnormal grain growth during homogenization. Meanwhile, the alloy with only 0.1Sc exhibited finer grains after FSP and T6 treatment than the alloys subjected to the same process but with higher Sc additive amount. Alloys with rolling-induced elongated grain structure exhibited better mechanical properties, and alloys with FSP-induced fine equiaxed grain structure exhibited higher high-strain and high-temperature internal friction values. These features are important performance parameters for applications in fields where vibration and noise are sensitive. The optimum additive amounts of Sc for alloys with elongated and fine equiaxed grain structures were 0.25 and 0.1, respectively.

[Complexity of Detecting CRISPR/Cas9-Mediated Homologous Recombination in Zebrafish].

Homology-directed (HD) genome modification offers an opportunity to precisely modify the genome. Despite reported successful cases, for many loci, precise genome editing remains challenging and inefficient in vivo. Here we report an effort to precisely knock-in a GFP reporter into gad locus mediated by CRISPR/Cas9 system in the zebrafish Danio rerio. PCR artifact was detected in testing for homologous recombination (HR), but was mitigated by optimizing PCR condition and decreasing the injected targeting plasmid concentration. Under this optimized condition, time course analysis revealed a decline of the HR-positive embryos at embryogenesis progressed. GFP signals also diminished at later developmental stages. The GFP signals were consistent with PCR detection, both of which suggested the loss of targeted insertion events at later stages. Such loss of insertion might be one underlying reason for the inability to obtain germ-line transgenic lines with GFP knocked into the gad locus. Our results suggest that the low HR efficiency associated with CRISPR-mediated knock-in is in part due to loss of insertion after targeted integration into the gad locus.