Analysis of the gut microbiota composition of myostatin mutant cattle prepared using CRISPR/Cas9

Abstract

Myostatin (MSTN) negatively regulates muscle development and positively regulates metabolism through various pathways. Although MSTN function in cattle has been widely studied, the changes in the gut microbiota due to MSTN mutation, which contribute to host health by regulating its metabolism, remain unclear. Here, high-throughput sequencing of the 16S rRNA gene was conducted to analyze the gut microbiota of wild-type (WT) and MSTN mutant (MT) cattle. A total of 925 operational taxonomic units (OTUs) were obtained, which were classified into 11 phyla and 168 genera. Alpha diversity results showed no significant differences between MT and WT cattle. Beta diversity analyses suggested that the microbial composition of WT and MT cattle was different. Three dominant phyla and 21 dominant genera were identified. The most abundant bacterial genus had a significant relationship with the host metabolism. Moreover, various bacteria beneficial for health were found in the intestines of MT cattle. Analysis of the correlation between dominant gut bacteria and serum metabolic factors affected by MSTN mutation indicated that MSTN mutation affected the metabolism mainly by three metabolism-related bacteria, Ruminococcaceae_UCG-013, Clostridium_sensu_stricto_1, and Ruminococcaceae_UCG-010. This study provides further insight into MSTN mutation regulating the host metabolism by gut microbes and provides evidence for the safety of gene-edited animals.

Gut fungal community composition analysis of myostatin mutant cattle prepared by CRISPR/Cas9

“…Additionally, intestinal fungi are closely related to bacteria, and their stable antagonistic and mutually beneficial symbiosis is fundamental to the homeostatic balance of intestinal microorganisms. The present study previously detected bacterial communities in the MSTN mutant cattle and found that MSTN mutations affected the metabolism of the host by affecting the composition of the bacteria (10). However, knowledge of the characteristics of the gut fungal community composition in the MSTN mutation cattle is still limited.…”

“…The cattle used in the present study were presumably 24-month-old females raised as common beef cattle in a local ranch. Sample collection was performed as described in an earlier study ( 10 ). Total DNA was extracted from fecal samples using the QlAamp Fast DNA Stool Mini Kit (Qiagen, Germany) according to the manufacturer's instructions.…”

“…Luxi cattle with the MSTN gene mutations prepared by the Inner Mongolia University through the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/CAS9) technology have more developed muscles and a higher lean meat yield ( 9 ). Previous studies had analyzed the changes in gut bacteria composition in MSTN -mutated cattle and found that the MSTN mutation affects the metabolism of the body by regulating the composition of gut bacteria, thereby affecting the growth traits of beef cattle ( 10 , 11 ). However, there are no reports on the changes in MSTN mutant bovine gut fungi and their correlation with host metabolism.…”

Inactivation of the MSTN gene expression changes the composition and function of the gut microbiome in sheep

“…These findings suggest that the gut microbiome has a synergistic effect on host growth and health. Previous studies have revealed a positive change of microbiota in pigs and cattle following mutation of the MSTN [ 20 , 21 ]. However, the effect of MSTN gene editing on the composition and function of gut microbiota in sheep is lacking.…”

Genetic polymorphism of myostatin gene in Sumba Ongole (Bos indicus) cattle and its association with growth traits

“…Skeletal muscle growth is controlled by myostatin ( MSTN ), sometimes called growth and differentiation factor 8 [ 1 – 4 ]. It has been mapped to bovine chromosome 2 and is composed of three exons and two introns [ 5 , 6 ].…”