经典基因工程和新的基因组编辑技术,特别是CRISPR/Cas技术,增加了改变生物体遗传物质的可能性。这些技术有潜力提供新的农业性状,包括改性微生物和环境应用。然而,合理的安全问题是由非预期的遗传修饰(GM)引起的,据报道这些技术会产生副作用.这里,我们系统地回顾了科学文献,研究了通过以下转基因技术修饰的植物中意外基因组改变的研究:根癌农杆菌介导的基因转移,生物轰击,和CRISPR-Cas9通过土壤杆菌介导的基因转移(基于DNA),生物弹轰击(基于DNA)和核糖核蛋白复合物(RNP)。我们的文献综述的结果表明,这些技术对宿主基因组的影响从小核苷酸多态性到大基因组变异,例如分段复制,染色体截断,三体,嗜铬细胞,断裂融合桥,包括DNA载体-骨架序列的大重排。我们还回顾了用于研究基因组改变的分析方法的类型,发现只有五篇文章在其分析方法中使用了全基因组测序。此外,如果没有长读数测序策略,一些研究中检测到的较大结构变异是不可能的,这表明文献中对这种影响的潜在低估。随着新技术的不断发展,未来应该对前瞻性分析方法进行更彻底的研究。这将为在转基因和基因编辑生物领域工作的监管机构提供有关检测和识别基因组干预措施能力的有价值的信息。
Classical genetic engineering and new genome editing techniques, especially the CRISPR/Cas technology, increase the possibilities for modifying the genetic material in organisms. These technologies have the potential to provide novel agricultural traits, including modified microorganisms and environmental applications. However, legitimate safety concerns arise from the unintended genetic modifications (GM) that have been reported as side-effects of such techniques. Here, we systematically
review the scientific literature for studies that have investigated unintended genomic alterations in plants modified by the following GM techniques: Agrobacterium tumefaciens-mediated gene transfer, biolistic bombardment, and CRISPR-Cas9 delivered via Agrobacterium-mediated gene transfer (DNA-based), biolistic bombardment (DNA-based) and as ribonucleoprotein complexes (RNPs). The results of our literature
review show that the impact of such techniques in host genomes varies from small nucleotide polymorphisms to large genomic variation, such as segmental duplication, chromosome truncation, trisomy, chromothripsis, breakage fusion bridge, including large rearrangements of DNA vector-backbone sequences. We have also reviewed the type of analytical method applied to investigate the genomic alterations and found that only five articles used whole genome sequencing in their analysis methods. In addition, larger structural variations detected in some studies would not be possible without long-read sequencing strategies, which shows a potential underestimation of such effects in the literature. As new technologies are constantly evolving, a more thorough examination of prospective analytical methods should be conducted in the future. This will provide regulators working in the field of genetically modified and gene-edited organisms with valuable information on the ability to detect and identify genomic interventions.