发明名称 |
MUTANT CAS9 PROTEINS |
摘要 |
Methods of making deletion mutants of Cas9 proteins and making chimeric Cas9 proteins are described. The Cas9 N- and C-terminal domains may play critical roles in crRNAitracrRNA binding and/or PAM selectivity. To analyze activity, a series of domain exchange mutants between NM and STI (Streptococcus thermophilus Cas9) were made, replacing the N and/or C terminus of NM (Neisseria meningitides Cas9) with the homologous region from STI. The chimeric proteins were then tested using the transcriptional reporter assay described herein altering the guideRNA and/or Cas9 specific PAM within the reporter to determine the influence of the domain exchanges on protein specificity. None of the N- terminal domain swaps between NM and STI endowed NM with novel properties. The C-terminal exchange generated a NM-STI hybrid that was capable of interacting with the STI crRNAitracrRNA complex and was further able to suppress a reporter with a STI specific PAM. |
申请公布号 |
CA2930829(A1) |
申请公布日期 |
2015.05.28 |
申请号 |
CA20142930829 |
申请日期 |
2014.11.19 |
申请人 |
PRESIDENT AND FELLOWS OF HARVARD COLLEGE |
发明人 |
CHAVEZ, ALEJANDRO;POELWIJK, FRANCK;CHURCH, GEORGE M. |
分类号 |
C12N15/00;C07H21/04;C07K1/00;C12N9/16 |
主分类号 |
C12N15/00 |
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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