Here, we efficiently generated transgenic cattle using two transposon systems (Sleeping Beauty and Piggybac) and their genomes were analyzed by next-generation sequencing (NGS). Blastocysts derived from microinjection of DNA transposons were selected and transferred into recipient cows. Nine transgenic cattle have been generated and grown-up to date without any health issues except two. Some of them expressed strong fluorescence and the transgene in the oocytes from a superovulating one were detected by PCR and sequencing. To investigate genomic variants by the transgene transposition, whole genomic DNA were analyzed by NGS. We found that preferred transposable integration (TA or TTAA) was identified in their genome. Even though multi-copies (i.e. fifteen) were confirmed, there was no significant difference in genome instabilities. In conclusion, we demonstrated that transgenic cattle using the DNA transposon system could be efficiently generated, and all those animals could be a valuable resource for agriculture and veterinary science.
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http://dx.doi.org/10.1038/srep27185 | DOI Listing |
JDS Commun
November 2024
Department of Preventive Veterinary Medicine and Animal Reproduction, School of Agricultural and Veterinary Sciences, São Paulo State University (UNESP), Jaboticabal, 14884-900, Brazil.
Genome editing is the latest breeding tool capable of accelerating the rate of genetic improvement for health and well-being traits in food animals. It enables the introduction of beneficial alleles within a single generation, including those that are of low frequency or absent in the population, while effectively bypassing linkage drag. For the dairy industry, genome editing can be used to make rapid genetic improvements that are precise, efficient, and transgene-free for functional traits that are not practically addressed without disrupting conventional breeding goals for overall economic merit based on genomic selection.
View Article and Find Full Text PDFParasit Vectors
October 2024
State Key Laboratory for Animal Disease Control and Prevention Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510642, China.
FEBS J
November 2024
Université Paris-Saclay, INRAE, UVSQ, VIM, Jouy-en-Josas, France.
Int J Mol Sci
August 2024
Key Laboratory of Livestock Biology, Northwest A&F University, Yangling 712100, China.
Prime editor, an editing tool based on the CRISPR/Cas9 system, allows for all 12 types of nucleotide exchanges and arbitrary indels in genomic sequences without the need for inducing DNA double-strand breaks. Despite its flexibility and precision, prime editing efficiency is still low and hindered by various factors such as target sites, editing types, and the length of the primer binding site. In this study, we developed a prime editing system by incorporating an RNA motif at the 3' terminal of the pegRNA and integrating all twin prime editor factors into a single plasmid.
View Article and Find Full Text PDFmBio
October 2024
Virology Division, USA Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Maryland, USA.
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