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acCRISPR:一种活动校正方法,用于提高CRISPR屏幕的准确性

Adithya Ramesh¹, Varun Trivedi¹, Sangcheon Lee¹

¹Department of Chemical and Environmental Engineering, University of California, Riverside, CA, 92521, USA.

Communications biology

|June 8, 2023

概括

我们开发了acCRISPR,这是一个计算工具,用于纠正CRISPR屏幕中的引导RNA效率. 这种方法准确地识别了必要的基因,改善了Yarrowia lipolytica等生物体的功能基因组学.

科学领域:

功能性基因组学是一种功能性基因组学.
基因编辑CRISPR-Cas9和CRISPR-Cas12a的基因编辑方法是不同的.
高通量选的高通量选

背景情况:

克里斯普尔屏幕对于理解基因功能至关重要,但受到可变导向RNA (sgRNA) 切割效率的挑战.
不有效的sgRNAs可以掩盖基因破坏的真实影响,使必要基因的识别复杂化.

研究的目的:

开发一个端到端的计算管道,acCRISPR,用于在聚合的CRISPR屏幕中准确识别必要的基因.
为纠正sgRNA切割效率的变化,从而提高选结果的可靠性.

主要方法:

acCRISPR从下一代测序数据中分析sgRNA读数.
它结合了经过实验确定的sgRNA切割效率,以调整查结果.
管道计算一个优化指标来确定基因适应效应.

主要成果:

在葡萄糖生长条件下,acCRISPR成功地在Yarrowia lipolytica中确定了必需基因.
该工具还用于识别Y. lipolytica.中与盐耐受性相关的基因.
确定了一组高可信度的基本基因,用于工业化油化学生产.

结论:

acCRISPR为基于CRISPR的功能基因组学提供了一个强大的实验计算框架.
这种方法提高了在聚合的CRISPR屏幕中基本基因识别的准确性.
该框架可适应在其他非传统生物体中使用.

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