4.5 Article

Engineering Cas9 for human genome editing

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CURRENT OPINION IN STRUCTURAL BIOLOGY
卷 69, 期 -, 页码 86-98

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CURRENT BIOLOGY LTD
DOI: 10.1016/j.sbi.2021.03.004

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Since the discovery of CRISPR-Cas9, labs worldwide have used this gene editing tool to modify the genomes of living cells. Scientists and engineers have altered Cas9 to make gene editing more precise and efficient. Key aspects of Cas9 engineering include sgRNA manipulation, PAM-recognition, specificity, deaminase fusions, reverse-transcriptase fusions, and structural rearrangements.
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.

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