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Efforts to Improve the Efficiency and Specificity of CRISPR-Cas9 Techniques

机译：努力提高CRISPR-CAS9技术的效率和特异性

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The CRISPR-Cas9 technique derived from bacterial immune system has been used in the majority of genome engineering applications due to its advantages of low cost and flexibility. In order to achieve efficient modifications to target genome in different cell types by CRISPR-Cas9 genome editing, three basic components are required, including 20nt sequence of sgRNA that could recognize desired DNA sequences, Cas9 protein for introducing double-strand breaks and the NGG PAM sequence for the binding between Cas9 proteins and the target genomic sites. However, the CRISPR-Cas9 technique today are facing some difficulties in adoption of further genome applications, especially those requiring high precision modifications. This is mainly because of the off-target effects, in which the spCas9 nuclease cleaves unwanted genomic sequences. To address this, a number of strategies have been developed. In this article, we mainly reviewed the factors in determining the activity of Cas9 proteins and concluded the approaches to reduce the off-target effects.
机译：由于其优点和灵活性低，因此在大多数基因组工程应用中，源自细菌免疫系统的CRISPR-CAS9技术已被用于大多数基因组工程应用。为了通过CRISPR-CAS9基因组编辑实现对不同细胞类型的靶基因组的有效修饰，需要三种碱性组分，包括20NT的SGRNA序列，可以识别所需的DNA序列，Cas9蛋白，用于引入双轴断裂和NGG PAM Cas9蛋白与靶基因组位点之间结合的顺序。然而，今天CRISPR-CAS9技术在采用进一步的基因组应用方面面临一些困难，尤其是需要高精度修改的困难。这主要是因为偏离目标效果，其中SPCAS9核酸酶切割不需要的基因组序列。为了解决这个问题，已经开发了许多策略。在本文中，我们主要综述了确定Cas9蛋白的活性的因素，并得出结论方法，以降低偏离目标效应。

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