This study successfully established a YE1-AncBE4max-based platform for efficient, precise dual-gene editing in goats and showed that the edited goats exhibited significantly increased body weight and a "double-muscling" phenotype by 90 days of age compared to wild-type controls.
The study successfully established CRISPR/Cas9 plasmid constructs for the mstnb gene in L. rohita and demonstrated their transfection in LRDM cell line across multiple passages, providing a basis for future studies on genome editing approaches using CRISPR/Cas9 constructs in fish muscle cell lines.
The feasibility of generating MSTN/BLG double gene-edited cattle embryos using a single CRISPR/EOCas12i plasmid and SCNT is demonstrated, providing a robust platform for multiplex genome editing aimed at improving meat production and milk traits.
Furui Wang, Lei Chen, Yuting Ning et al.· Frontiers in Genome Editing· 0 citations
The effects of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9‐mediated disruption of the myostatin gene (mstnb1) on the growth performance of rainbow trout (Oncorhynchus mykiss) were evaluated. Two specific guide RNAs (gRNAs) targeting the first exon of the target gene were co‐injected into fertilized eggs. While the survival rate of the injected embryos at the swim‐up stage was 23.5%, Sanger sequencing of F0 individuals confirmed the successful generation of mutations. During a 10‐month growing period, quantifiable phenotypic data revealed a significant enhancement in macroscopic somatic growth. The mstnb1‐F0 mosaic fish exhibited an approximate 56% increase in body weight compared to the wild‐type (WT) control group. Specifically, the mutant group reached an average weight of 825.0 ± 12.3 g, significantly outperforming the WT group, which reached 464.1 ± 9.2 g. Our results demonstrate that disruption of the mstnb1 gene using CRISPR/Cas9 technology induces a striking somatic weight gain rather than merely altering microscopic muscle fiber characteristics, demonstrating high biotechnological potential for shortening production time and reducing unit costs in sustainable rainbow trout aquaculture.
A. Bayır, M. Bayır, Wenjing Tao et al.· Journal of the World Aquacul...· 0 citations
The functions of noncoding variants associated with complex traits in livestock remain poorly understood. In this study, we investigated two candidate noncoding variants within the XKR4-CHCHD7 locus identified from our previous analysis. Dual-luciferase reporter assays demonstrated allele-specific regulatory activity of these two regions in bovine muscle satellite cells (BMSCs), 293T cells, and C2C12 cells. Endogenous deletion of the candidate regions using a clustered regularly interspaced short palindromic repeats (CRISPR)-based high-fidelity Cas12Max (hfCas12Max) system revealed that the region containing chr14:22840845 (SNP-0845) exerted broader effects on BMSC function including reduced proliferation and migration, altered cell-cycle progression, and enhanced myogenic differentiation. Expression screening of candidate effector genes further identified PENK and TMEM68 as downstream candidate genes for SNP-0845. Rescue experiments further showed that PENK exerted stronger recovery effects than TMEM68 on the proliferation and migration defects caused by deletion of this region, supporting PENK as a major candidate effector downstream of the SNP-0845. Functional assays showed that PENK knockdown impaired BMSC proliferation and migration while promoting myogenic differentiation, whereas PENK overexpression partially reversed these effects. In vivo Penk knockdown reduced quadriceps femoris weight and altered muscle fiber composition in mice. Collectively, our findings suggest that a noncoding regulatory region modulates BMSC fate and muscle growth-related processes through PENK.
Tianyi Wu, Feng Liu, Qunhao Niu et al.· International Journal of Mol...· 0 citations
Findings reveal a novel epigenetic regulatory axis (novel-miR-357/S100A2 mRNA) involved in early muscle development and provide an important molecular mechanism for deeper understanding of the epigenetic regulatory processes underlying heterosis in cattle.
Gaoqing Xu, Chunqi Hou, He Ding et al.· Animal Genetics· 0 citations
The phenotypic profile of a CRISPR-generated Ent2 mutant line is described and the feasibility of combining genome editing with balancer chromosome strategies in Drosophila is demonstrated, demonstrating the feasibility of combining genome editing with balancer chromosome strategies in Drosophila.
Chikun Li, Lin Li, Ying Chen et al.· Journal of Visualized Experi...· 0 citations
