The food safety assessment of all-female common carp (Cyprinus carpio) (cyp17a1+/-;XX genotype) generated using genome editing technology.

There are several technical challenges and public issues concerning genome editing applications before they become viable in commercial aquaculture. Recently, we developed a novel strategy to generate all-female (AF) common carp, which exhibited a growth advantage over the control carp, using genetic editing through single gene-targeting manipulation. Here, we found that the body weight of the AF common carp was higher by 22.58% than that of the control common carp. Because the genotype of the AF common carp was cyp17a1+/-;XX, the contents of sex steroids were normally synthesized, as they were comparable to that of the control female carp. To evaluate the food safety of the AF carp, Wistar rats were fed a diet containing control female carp (control, C) or all-female (AF) carp at an incorporation rate of 5, 10 and 20% (w/w) for 90 days. Compared with those fed control carp, the rats fed AF common carp exhibited no significant difference in body weight, food intake, feed conversion ratio, hematology, serum biochemistry, urine test, relative organ weight, gross necropsy, and histopathological examination. This is the first food safety assessment of the farmed fish strain cultured using CRISPR/Cas9, which will further advance the fishery development of genome-edited animals.

siRNA-Mediated MrIAG Silencing Induces Sex Reversal in Macrobrachium rosenbergii.

The insulin-like androgenic gland (IAG) gene is well known in male crustacean, and it is a key regulator in male sexual differentiation and maintaining the male sexual characteristic. The neo-female can be produced by silencing the MrIAG (Macrobrachium rosenbergii Insulin-like Androgenic Gland) in male Macrobrachium rosenbergii. This is the first time to use siRNA approach to silenced MrIAG in male M. rosenbergii. In the current study, the optimal injection dosage to achieve sex reversal is 0.5 µg/g body weight. After MrIAG silencing, the expression level of Dmrt11e, Dmrt99b, MRPINK, Mrr, Sxl1, and Sxl2 decreased significantly. As their long-term silencing effect of MrIAG, the dsRNA and siRNA approaches produce three and two individual neo-females, respectively. The neo-female has a wider brood chamber, ovipositing setae, and ovigerous setae, which is resembled normal female. After a long-term silencing with siRNA, most of the germ cells were arrested in spermatocytes stage, but the spermatocytes in control can further developed into spermatozoon. The seminiferous tubules are loosely arranged and the spermatocytes are more than spermatozoon in the 0.5 µg/g body weight treatment dose. This current study suggests a new path to obtain neo-females through siRNA silencing.