Biosystems and Biomaterials Division, Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States.
Biochemistry. 2023 Dec 19;62(24):3500-3511. doi: 10.1021/acs.biochem.2c00431. Epub 2022 Oct 28.
Genome editing is a rapidly evolving biotechnology with the potential to transform many sectors of industry such as agriculture, biomanufacturing, and medicine. This technology is enabled by an ever-growing portfolio of biomolecular reagents that span the central dogma, from DNA to RNA to protein. In this paper, we draw from our unique perspective as the National Metrology Institute of the United States to bring attention to the importance of understanding and reporting genome editing formulations accurately and promoting concepts to verify successful delivery into cells. Achieving the correct understanding may be hindered by the way units, quantities, and stoichiometries are reported in the field. We highlight the variability in how editing formulations are reported in the literature and examine how a reference molecule could be used to verify the delivery of a reagent into cells. We provide recommendations on how more accurate reporting of editing formulations and more careful verification of the steps in an editing experiment can help set baseline expectations of reagent performance, toward the aim of enabling genome editing studies to be more reproducible. We conclude with a future outlook on technologies that can further our control and enable our understanding of genome editing outcomes at the single-cell level.
基因组编辑是一种快速发展的生物技术，有可能改变农业、生物制造和医学等许多行业。这项技术得益于越来越多的生物分子试剂，这些试剂跨越了从 DNA 到 RNA 再到蛋白质的中心法则。在本文中，我们从美国国家计量研究所的独特视角出发，提请人们注意准确理解和报告基因组编辑配方的重要性，并推广验证成功递送到细胞的概念。单位、数量和化学计量的报告方式可能会阻碍对正确理解的达成。我们强调了文献中编辑配方报告的可变性，并研究了如何使用参考分子来验证试剂递送到细胞中。我们就如何更准确地报告编辑配方以及更仔细地验证编辑实验中的步骤提出了建议，这有助于设定试剂性能的基线预期，以实现基因组编辑研究的更高可重复性。最后，我们展望了未来的技术，这些技术可以进一步控制并使我们能够在单细胞水平上理解基因组编辑的结果。
