Institute of Biotechnology, Vilnius University, Saulėtekio av. 7, LT-10257 Vilnius, Lithuania.
Institute of Biotechnology, Vilnius University, Saulėtekio av. 7, LT-10257 Vilnius, Lithuania.
Curr Opin Microbiol. 2017 Jun;37:88-94. doi: 10.1016/j.mib.2017.05.009. Epub 2017 Jun 20.
In the past few years, the Cas9 endonuclease from the type II CRISPR-Cas bacterial antiviral defense system has revolutionized the genome editing field. Guided by an RNA molecule, Cas9 can be reprogrammed to target almost any DNA sequence: the only limitation being the short nucleotide sequence in the vicinity of the target, termed the PAM, which is characteristic for each Cas9 protein. Streptococcus pyogenes Cas9 which recognizes the NGG PAM is currently most widely used for genome manipulation. However, Cas9 orthologues and engineered Cas9 variants offer expanded genome targeting capabilities, improved specificity and biochemical properties.
在过去的几年中，来自 II 型 CRISPR-Cas 细菌抗病毒防御系统的 Cas9 内切酶彻底改变了基因组编辑领域。在 RNA 分子的引导下，Cas9 可以被重新编程以靶向几乎任何 DNA 序列：唯一的限制是靶标附近的短核苷酸序列，称为 PAM，这是每种 Cas9 蛋白的特征。目前，最常用于基因组操作的是识别 NGG PAM 的化脓性链球菌 Cas9。然而，Cas9 同源物和工程化的 Cas9 变体提供了扩展的基因组靶向能力、提高的特异性和生化特性。
