Bacterial and virus strains |
E. coli Bl21(DE3) competent cells |
NEB |
Cat# C2527I |
E. coli DH5a competent cells |
NEB |
Cat# 18265017 |
|
|
|
Chemicals, peptides, and recombinant proteins |
Modified and unmodified oligos |
IDT |
N/A |
IPTG |
GoldBio |
Cat # I2481C500 |
LB broth |
Teknova |
#L9145 |
Restriction enzymes |
NEB |
Choose from catalog |
iProof high fidelity PCR kit |
Biorad |
#1725331 |
Acrylamide |
thermal |
HC2040 |
Urea |
VWR |
57-13-6 |
Fluorescence stain Cy3 and Cy5 |
Lumiprobe |
11320 and 13320 |
SYPRO® Orange |
Thermo Fisher |
S6650 |
Formamide |
VWR |
75-12-7 |
MnCl2
|
Fisher Scientific |
7773-01-5 |
Strep resin |
IBA |
2-1201-025 |
Nickel resin |
QIAGEN |
Cat. No. / ID: 30250 |
lateral flow strip |
HybriDetect 1 |
MGDS |
Copper Grids 1.2/1.3 200 mesh |
QUANTIFOIL |
Q250-CR1.3 |
Type I-A Cascade |
This study |
N/A |
Cas3 HEL |
This study |
N/A |
Cas3 HD |
This study |
N/A |
T.fusca type I-E Cascade |
Xiao et al., 2017
|
N/A |
T.fusca type I-E Cas3 |
Xiao et al., 2018
|
N/A |
|
|
|
Deposited data |
Cascade alone |
This study |
PDB: 7TR6 and EMBD-26081 |
Cascade-Cas3 |
This study |
PDB: 7TR8 and EMBD-26082 |
Partial R-loop |
This study |
PDB: 7TR9 and EMBD-26083 |
Full R-loop |
This study |
PDB: 7TRA and EMBD-26084 |
Cas3-Cas8a alone |
This study |
EMBD-26097 |
Experimental models: Cell lines |
HAP1_GFP |
Dolan et al. 2019
|
N/A |
HEK293_GFP |
This study |
N/A |
Recombinant DNA |
Complete list of plasmids |
This study |
Table S2
|
Oligonucleotides |
Complete list of DNA oligonucleotide sequences |
This study |
Table S3
|
Miniature CRISPR array spacers |
This study |
Table S3
|
Software and algorithms |
EPU |
Thermo Fisher Scientific |
N/A |
RELION 3 |
Scheres Lab |
https://www3.mrc-lmb.cam.ac.uk/relion/index.php?title%20=%20Main_PAGE
|
cryoSPARC |
Ali Punjani et al., 2017
|
https://cryosparc.com/
|
Coot (v. 0.9.5.1-pre) |
(Emsley and Cowtan, 2004; Emsley et al., 2010) |
https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
|
PHENIX |
Adams et al. (2010) |
https://phenix-online.org/
|
ChimeraX (v. 1.2) |
(Goddard et al., 2018) |
https://www.cgl.ucsf.edu/chimerax/
|
ClustalW |
(Goujon et al., 2010; Sievers et al., 2011) |
https://www.ebi.ac.uk/Tools/msa/clustalo/
|
Other |
MonoQ 5/50 GL |
Cytiva |
Cat# 17516601 |
HiLoad 16/600 Superdex 200 pg |
Cytiva |
Cat# 28989335 |
Quantifoil R 1.2/1.3, 200 mesh, copper |
Quantifoil |
Q250-CR1.3 |