Beam Therapeutics, 26 Landsdowne St., Cambridge, MA 02139, USA.
Curr Opin Struct Biol. 2021 Aug;69:86-98. doi: 10.1016/j.sbi.2021.03.004. Epub 2021 May 5.
Since the initial reports describing CRISPR-Cas9, labs across the globe have leveraged this valuable gene editing tool to alter the genomes of living cells. With the goal of generating more precise and efficient genome changes, scientists and engineers have mutated, evolved, and covalently altered Cas9 in order to predictably edit the genetic code. Here, we highlight recent advancements and contributions to the growing field of Cas9 engineering. We present key aspects of Cas9 engineering efforts focused on sgRNA manipulation, PAM-recognition, specificity, deaminase fusions, reverse-transcriptase fusions, and structural rearrangements of this important gene-modifying tool.
自最初描述 CRISPR-Cas9 的报告发布以来，全球各地的实验室都利用这一宝贵的基因编辑工具来改变活细胞的基因组。为了产生更精确和高效的基因组变化，科学家和工程师已经对 Cas9 进行了突变、进化和共价修饰，以便对遗传密码进行可预测的编辑。在这里，我们重点介绍 Cas9 工程领域的最新进展和贡献。我们介绍了 Cas9 工程努力的关键方面，重点是 sgRNA 的操作、PAM 的识别、特异性、脱氨酶融合、逆转录酶融合以及这个重要基因修饰工具的结构重排。
