RT Journal Article
SR Electronic
T1 Direct purification of CRISPR/Cas ribonucleoprotein from <em>E. coli</em> in a single step
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 377366
DO 10.1101/377366
A1 Siyu Lin
A1 Jie Qiao
A1 Lixin Ma
A1 Yi Liu
YR 2018
UL http://biorxiv.org/content/early/2018/07/26/377366.abstract
AB CRISPR/Cas ribonucleoprotein (RNP) complexes have been recently used as promising biological tools with plenty of applications, however, there are by far no efficient methods to prepare them at large scale and low cost. Here, we present a simple method to directly produce and purify Cas RNP, including the widely used Cas9 and Cas12a nuclease, from E.coli in a single step using an ultra-high-affinity CL7/Im7 purification system. The prepared Cas RNP shows high stability, solid nuclease activity in vitro, and profound genome editing efficiency in vivo. Our method is convenient, cost-effective, and applicable to prepare other CRISPR associated nucleases.