In:
eLife, eLife Sciences Publications, Ltd, Vol. 8 ( 2019-08-09)
Abstract:
CRISPR-Cas systems provide bacteria and archaea with programmable immunity against mobile genetic elements. Evolutionary pressure by CRISPR-Cas has driven bacteriophage to evolve small protein inhibitors, anti-CRISPRs (Acrs), that block Cas enzyme function by wide-ranging mechanisms. We show here that the inhibitor AcrVA4 uses a previously undescribed strategy to recognize the L. bacterium Cas12a (LbCas12a) pre-crRNA processing nuclease, forming a Cas12a dimer, and allosterically inhibiting DNA binding. The Ac. species Cas12a (AsCas12a) enzyme, widely used for genome editing applications, contains an ancestral helical bundle that blocks AcrVA4 binding and allows it to escape anti-CRISPR recognition. Using biochemical, microbiological, and human cell editing experiments, we show that Cas12a orthologs can be rendered either sensitive or resistant to AcrVA4 through rational structural engineering informed by evolution. Together, these findings explain a new mode of CRISPR-Cas inhibition and illustrate how structural variability in Cas effectors can drive opportunistic co-evolution of inhibitors by bacteriophage.
Type of Medium:
Online Resource
ISSN:
2050-084X
DOI:
10.7554/eLife.49110.001
DOI:
10.7554/eLife.49110.002
DOI:
10.7554/eLife.49110.003
DOI:
10.7554/eLife.49110.004
DOI:
10.7554/eLife.49110.005
DOI:
10.7554/eLife.49110.006
DOI:
10.7554/eLife.49110.007
DOI:
10.7554/eLife.49110.008
DOI:
10.7554/eLife.49110.009
DOI:
10.7554/eLife.49110.010
DOI:
10.7554/eLife.49110.011
DOI:
10.7554/eLife.49110.012
DOI:
10.7554/eLife.49110.013
DOI:
10.7554/eLife.49110.014
DOI:
10.7554/eLife.49110.015
DOI:
10.7554/eLife.49110.016
DOI:
10.7554/eLife.49110.017
DOI:
10.7554/eLife.49110.018
DOI:
10.7554/eLife.49110.019
DOI:
10.7554/eLife.49110.020
DOI:
10.7554/eLife.49110.021
DOI:
10.7554/eLife.49110.022
DOI:
10.7554/eLife.49110.023
DOI:
10.7554/eLife.49110.024
DOI:
10.7554/eLife.49110.025
DOI:
10.7554/eLife.49110.026
DOI:
10.7554/eLife.49110.027
DOI:
10.7554/eLife.49110.028
DOI:
10.7554/eLife.49110.029
DOI:
10.7554/eLife.49110.030
DOI:
10.7554/eLife.49110.036
DOI:
10.7554/eLife.49110.037
Language:
English
Publisher:
eLife Sciences Publications, Ltd
Publication Date:
2019
detail.hit.zdb_id:
2687154-3