Heritable Targeted Inactivation of Myostatin Gene in Yellow Catfish (Pelteobagrus fulvidraco) Using Engineered Zinc Finger Nucleases

Abstract

Yellow catfish (Pelteobagrus fulvidraco) is one of the most important freshwater aquaculture species in China. However, its small size and lower meat yield limit its edible value. Myostatin (MSTN) is a negative regulator of mammalian muscle growth. But, the function of Mstn in fish remains elusive. To explore roles of mstn gene in fish growth and create a strain of yellow catfish with high amount of muscle mass, we performed targeted disruption of mstn in yellow catfish using engineered zinc-finger nucleases (ZFNs). Employing zebrafish embryos as a screening system to identify ZFN activity, we obtained one pair of ZFNs that can edit mstn in yellow catfish genome. Using the ZFNs, we successfully obtained two founders (Founder July29-7 and Founder July29-8) carrying mutated mstn gene in their germ cells. The mutated mstn allele inherited from Founder July29-7 was a null allele (mstnnju6) containing a 4 bp insertion, predicted to encode function null Mstn. The mutated mstn inherited from Founder July29-8 was a complex type of mutation (mstnnju7), predicted to encode a protein lacking two amino acids in the N-terminal secretory signal of Mstn. Totally, we obtained 6 mstnnju6/+ and 14 mstnnju7/+ yellow catfish. To our best knowledge, this is the first endogenous gene knockout in aquaculture fish. Our result will help in understanding the roles of mstn gene in fish.

Zebrafish foxc1a Plays a Crucial Role in Early Somitogenesis by Restricting the Expression of aldh1a2 Directly

“…The method used to examine TALEN activity in zebrafish embryos was the same as described previously (21). The primers for amplifying the foxc1a fragment containing the potential TALEN targeting site were CCGTTTTGGA-GAGCAGTCA (forward) and GTCTCCGGCCTGGTTCAG (reverse).…”

Genome Editing with Engineered Nucleases in Economically Important Animals and Plants: State of the Art in the Research Pipeline

“…Apart from livestock species, SDNs were also used in fish. Using ZFNs, Dong et al (2011) successfully mutated the mstn gene in yellow catfish (Pelteobagrus fulvidraco), an important freshwater aquaculture species.…”

Generation of Myostatin b Knockout Yellow Catfish (Tachysurus Fulvidraco) Using Transcription Activator-Like Effector Nucleases

“…24 For each mating, eggs were collected by massaging a female yellow catfish in the abdomen repeatedly from anterior to posterior and were mixed with minced testes harvested from a male fish in 5 mL of 0.69% NaCl solution. The fertilized eggs were then dispensed into a dish with 30 mL of fresh water at a density of about 300 eggs per dish.…”

“…[21][22][23] Previously, we knocked out one of the duplicated copies of myostatin gene (mstna, previously named mstn) in yellow catfish (Tachysurus fulvidraco, previously named Pelteobagrus fulvidraco), one of the most important freshwater aquaculture species in China, using ZFN. 24 In this study, we report the identification of a second myostatin gene (mstnb) in yellow catfish. Using TALEN technology, we generated, from two founder fish, 13 heritable yellow catfish carrying either of the two function null alleles of mstnb.…”