Nature Communications ( IF 14.7 ) Pub Date : 2023-03-30 , DOI: 10.1038/s41467-023-37508-7
Nana Yan 1 , Hu Feng 1 , Yongsen Sun 1 , Ying Xin 1, 2 , Haihang Zhang 1 , Hongjiang Lu 1, 2 , Jitan Zheng 1, 3 , Chenfei He 1 , Zhenrui Zuo 1 , Tanglong Yuan 1 , Nana Li 1, 2 , Long Xie 1 , Wu Wei 4, 5 , Yidi Sun 6 , Erwei Zuo 1
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Base editors have been reported to induce off-target mutations in cultured cells, mouse embryos and rice, but their long-term effects in vivo remain unknown. Here, we develop a Systematic evaluation Approach For gene Editing tools by Transgenic mIce (SAFETI), and evaluate the off-target effects of BE3, high fidelity version of CBE (YE1-BE3-FNLS) and ABE (ABE7.10F148A) in ~400 transgenic mice over 15 months. Whole-genome sequence analysis reveals BE3 expression generated de novo mutations in the offspring of transgenic mice. RNA-seq analysis reveals both BE3 and YE1-BE3-FNLS induce transcriptome-wide SNVs, and the numbers of RNA SNVs are positively correlated with CBE expression levels across various tissues. By contrast, ABE7.10F148A shows no detectable off-target DNA or RNA SNVs. Notably, we observe abnormal phenotypes including obesity and developmental delay in mice with permanent genomic BE3 overexpression during long-time monitoring, elucidating a potentially overlooked aspect of side effects of BE3 in vivo.
中文翻译:
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胞嘧啶碱基编辑器在转基因小鼠中诱导脱靶突变和不良表型效应
据报道,碱基编辑器可在培养细胞、小鼠胚胎和水稻中诱导脱靶突变,但其在体内的长期影响仍不清楚。在这里,我们开发了 Transgenic mIce (SAFETI) 基因编辑工具的系统评估方法,并评估了 BE3、高保真版本 CBE (YE1-BE3-FNLS) 和 ABE (ABE7.10 F148A ) 在15 个月内约 400 只转基因小鼠。全基因组序列分析揭示了 BE3 表达在转基因小鼠的后代中产生了从头突变。 RNA-seq分析表明BE3和YE1-BE3-FNLS均诱导转录组范围的SNV,并且RNA SNV的数量与不同组织中的CBE表达水平呈正相关。相比之下,ABE7.10 F148A未显示出可检测到的脱靶 DNA 或 RNA SNV。值得注意的是,我们在长期监测过程中观察到了永久性基因组BE3过度表达的小鼠的异常表型,包括肥胖和发育迟缓,阐明了BE3体内副作用的一个可能被忽视的方面。