Multiplexed orthogonal genome editing and transcriptional activation by Cas12a
Multiplexed orthogonal genome editing and transcriptional activation by Cas12a
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DOI：
10.1038/s41592-018-0262-1
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发表时间：
2019-01-01
期刊：
影响因子：
48
通讯作者：
Tschaharganeh, Darjus F.
中科院分区：
文献类型：
--
作者：
Breinig, Marco;Schweitzer, Anabel Y.;Tschaharganeh, Darjus F.
用于正交敲除和基因激活的基于CRISPR-Cas9的组合扰动方法已经受到复杂载体设计和多个构建体的共递送的阻碍。在这里，我们证明了与转录激活因子结构域融合的催化活性CRISPR-Cas 12 a能够通过改变向导长度在基因组编辑和转录激活之间灵活切换。通过利用Cas 12 a介导的CRISPR-RNA阵列处理，我们说明了Cas 12 a-VPR能够在体外和体内简化多重敲除和转录激活。
CRISPR-Cas9-based combinatorial perturbation approaches for orthogonal knockout and gene activation have been impeded by complex vector designs and co-delivery of multiple constructs. Here, we demonstrate that catalytically active CRISPR-Cas12a fused to a transcriptional-activator domain enables flexible switching between genome editing and transcriptional activation by altering guide length. By leveraging Cas12a-mediated CRISPR-RNA array processing, we illustrate that Cas12a-VPR enables simplified multiplexed knockout and transcriptional activation in vitro and in vivo.