深度学习和 CRISPR-Cas13d 同源物发现,优化 RNA 靶向

IF 9 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Jingyi Wei, Peter Lotfy, Kian Faizi, Sara Baungaard, Emily Gibson, Eleanor Wang, Hannah Slabodkin, Emily Kinnaman, Sita Chandrasekaran, Hugo Kitano, Matthew G. Durrant, Connor V. Duffy, April Pawluk, Patrick D. Hsu, Silvana Konermann
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引用次数: 0

摘要

需要有效而精确的哺乳动物转录组工程技术来加速生物发现和 RNA 治疗。尽管可编程CRISPR-Cas13核糖核酸酶大有可为,但由于对引导RNA设计规则的不完全了解以及RNA脱靶或附带裂解造成的细胞毒性,它们的应用一直受到阻碍。在这里,我们量化了超过 127,000 条 RfxCas13d (CasRx) 引导 RNA 的性能,并系统地评估了七个机器学习模型,从而建立了一种引导效率预测算法,并在多种人类细胞类型中进行了正交验证。深度学习模型解释揭示了高效导引的首选序列主题和次要特征。我们接下来鉴定并筛选了46个新型Cas13d直向同源物,发现DjCas13d具有低细胞毒性和高特异性--即使在靶向敏感细胞类型(包括干细胞和神经元)中的丰富转录本时也是如此。我们的Cas13d引导效率模型成功地推广到了DjCas13d上,这说明了机器学习与直向同源物发现的结合在促进人类细胞RNA靶向方面的强大作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Deep learning and CRISPR-Cas13d ortholog discovery for optimized RNA targeting

Deep learning and CRISPR-Cas13d ortholog discovery for optimized RNA targeting

Effective and precise mammalian transcriptome engineering technologies are needed to accelerate biological discovery and RNA therapeutics. Despite the promise of programmable CRISPR-Cas13 ribonucleases, their utility has been hampered by an incomplete understanding of guide RNA design rules and cellular toxicity resulting from off-target or collateral RNA cleavage. Here, we quantified the performance of over 127,000 RfxCas13d (CasRx) guide RNAs and systematically evaluated seven machine learning models to build a guide efficiency prediction algorithm orthogonally validated across multiple human cell types. Deep learning model interpretation revealed preferred sequence motifs and secondary features for highly efficient guides. We next identified and screened 46 novel Cas13d orthologs, finding that DjCas13d achieves low cellular toxicity and high specificity—even when targeting abundant transcripts in sensitive cell types, including stem cells and neurons. Our Cas13d guide efficiency model was successfully generalized to DjCas13d, illustrating the power of combining machine learning with ortholog discovery to advance RNA targeting in human cells.

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来源期刊
Cell Systems
Cell Systems Medicine-Pathology and Forensic Medicine
CiteScore
16.50
自引率
1.10%
发文量
84
审稿时长
42 days
期刊介绍: In 2015, Cell Systems was founded as a platform within Cell Press to showcase innovative research in systems biology. Our primary goal is to investigate complex biological phenomena that cannot be simply explained by basic mathematical principles. While the physical sciences have long successfully tackled such challenges, we have discovered that our most impactful publications often employ quantitative, inference-based methodologies borrowed from the fields of physics, engineering, mathematics, and computer science. We are committed to providing a home for elegant research that addresses fundamental questions in systems biology.
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