Bioscience Methods 2025, Vol.16, No.2, 83-99 http://bioscipublisher.com/index.php/bm 97 we can test various hypotheses on the computer, such as "what happens if the activity of gene X is increased by 10%" and "what happens if the function of gene Y is deleted", and select the best improvement plan from them, and then verify it in real animals. This can save a lot of time and cost, and avoid unnecessary use of animals caused by blind trial and error. 7.3 Ethical, technical and practical considerations On the road to improving livestock using single-cell multi-omics and genetic engineering, we must also face some ethical, technical and practical challenges. Gene editing of animals and in-depth analysis of animal embryonic development may cause public concerns about animal welfare and biosafety. The general public may question whether such in-depth cell manipulation of animals will cause pain or unnatural changes to animals. Therefore, when promoting these studies and applications, we need to be transparent and open, actively carry out popular science communication, clarify the purpose and safety of the technology, and let the public understand that improving animal production performance does not mean "abuse" or "monsterization" of animals. The animals themselves will not suffer as a result, but may be healthier (such as reducing diseases or growth stagnation). At the same time, we must respect certain ethical bottom lines, such as not blindly breeding animals with high artificial intervention in large quantities in the natural environment to avoid impacting the ecology. Single-cell multi-omics still has room for improvement. As mentioned above, there are problems such as cost and high data complexity. This requires us to establish a cross-disciplinary cooperation team to introduce the latest sequencing technology and analysis algorithms into animal husbandry research. Interdisciplinary training is also necessary, so that traditional breeding experts can learn computational and molecular tools, and data scientists can understand biological problems. Technical details such as how to separate high-quality nuclei from meat livestock muscles with high fibroblast content, how to ensure the activity of samples transported over long distances, and how to deal with sample heterogeneity (such as batch effects of different individuals) need to be explored and solved. Perhaps several regional single-cell sequencing centers can be established to serve surrounding agricultural research units, focus on solving technical problems, and provide standardized services. In addition to the technology itself, the acceptance of farmers, economic benefits, and regulatory policies should also be considered when applying scientific research results to the breeding industry. New molecular breeding technologies may require certain investments, training, and strategic planning. For example, single-cell multi-omics-assisted breeding can be piloted in national core breeding farms first, and then gradually promoted after significant results are achieved. At the same time, corresponding industry standards and regulatory frameworks should be formulated, such as the identification, detection methods, and risk assessment systems of gene-edited animals. At present, the international regulation of gene editing of livestock and poultry is different. The United States has conditionally released related products from Japan and China, and they are also under discussion. We need to improve policies based on scientific evidence to encourage innovation and ensure safety. There are also issues of data ownership and privacy. Although animals do not have the concept of personal privacy like humans, breed resources belong to national or corporate assets. How to share and protect large-scale animal genetic and epigenetic data requires policy guidance. Perhaps a national livestock and poultry single-cell genomics database can be established, with data uploaded by various scientific research units, managed in a unified manner, and open for use in a graded manner. This can not only accelerate research progress, but also avoid data fragmentation and duplication of work. Finally, animal welfare must be mentioned. Any biotechnology method should be implemented without reducing animal welfare, and even with the goal of improving animal health and well-being. For example, if genetic improvement can make animals more resistant to disease and less prone to illness, this is a good thing for the animals themselves and should be strongly promoted. On the contrary, if a certain improvement leads to an increase in the physiological burden of the animal (such as extreme muscle development leading to skeletal and cardiac load), it needs to be treated with caution and production should not be sacrificed at the expense of animal health. Acknowledgments The authors thank the two anonymous peer reviewers for their thorough review of this study and for their valuable suggestions for improvement.
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