发明名称 |
CAS9-DNA TARGETING UNIT CHIMERAS |
摘要 |
The present invention provides a Cas9 platform to facilitate single-site nuclease gene editing precision within a human genome. For example, a Cas9 nuclease/DN A- targeting unit (Cas9-DTU) fusion protein precisely delivers a Cas9/sgRNA complex to a specific target site within the genome for subsequent sgRNA-dependent cleavage of an adjacent target sequence. Alternatively, attenuating Cas9 binding using mutations to the a protospacer adjacent motif (PAM) recognition domain makes Cas9 target site recognition dependent on the associated DTU, all while retaining Cas9's sgRNA-mediated DNA cleavage fidelity. Cas9-DTU fusion proteins have improved target site binding precision, greater nuclease activity, and a broader sequence targeting range than standard Cas9 systems. Existing Cas9 or sgRNA variants (e.g., truncated sgRNAs (tru-gRNAs), nickases and Fokl fusions) are compatible with these improvements to further reduce off-target cleavage. A robust, broadly applicable strategy is disclosed to impart Cas9 genome-editing systems with the single-genomic-site accuracy needed for safe, effective clinical application. |
申请公布号 |
WO2016106338(A2) |
申请公布日期 |
2016.06.30 |
申请号 |
WO2015US67424 |
申请日期 |
2015.12.22 |
申请人 |
UNIVERSITY OF MASSACHUSETTS |
发明人 |
WOLFE, SCOT, ANDREW;BOLUKBASI, MEHMET, FATIH;GUPTA, ANKIT;SONTHEIMER, ERIK, J.;AMRANI, NADIA |
分类号 |
C12N9/22 |
主分类号 |
C12N9/22 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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