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Adaptation by Type V-A and V-B CRISPR-Cas Systems Demonstrates Conserved Protospacer Selection Mechanisms Between Diverse CRISPR-Cas Types.

Wu WY, Jackson SA, Almendros C, Haagsma AC, Yilmaz S, Gort G, van der Oost J, Brouns SJJ, Staals RHJ
CRISPR J. 2022 Aug;5(4):536-547. doi: 10.1089/crispr.2021.0150. Epub 2022 Jul 12. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
186438pCas4_1_2_VAEncodes Cas4, Cas1, Cas2, leader, repeat and one spacer from F. Novicida (type V-A)
186439pCas4_1_Δ2_VAEncodes Cas4, Cas1, leader, repeat and one spacer from F. Novicida (type V-A)
186440pCasΔ4_1_2_VAEncodes Cas1, Cas2, leader, repeat and one spacer from F. Novicida (type V-A)
186441pCas-mut4_1_2_VAEncodes Cas4 (K70A), Cas1, Cas2, leader, repeat and one spacer from F. Novicida (type V-A)
186442pCas-4/1_2_VBEncodes Cas4/1, Cas2, leader, repeat and one spacer from A. acidoterrestris (type V-B)
186443pCas-4/1Δ2_VBEncodes Cas4/1, leader, repeat and one spacer from A. acidoterrestris (type V-B)
186444pCas-mut4/1_2_VBEncodes Cas4(K81A)/1, Cas2, leader, repeat and one spacer from A. acidoterrestris (type V-B)
186445pCas_4(I-U)/1_2_VBEncodes Cas4/1, Cas2, leader, repeat and one spacer from A. acidoterrestris (type V-B). Cas4 domain swapped with type U-I Cas4 domain from G. sulfurreducens
186446pCas12aEncodes Cas12a from F. novicida (type V-A)
186447pdCas12a(RuvC)Encodes catalytically inactive Cas12a (D917A, E1006A) from F. novicida (type V-A)
186448pCas12(PI)Encodes inactive Cas12a containing mutation in the PI domain (K613A, K617A) from F. novicida (type V-A)
186449pCas12bEncodes Cas12b from A. acidoterrestris (type V-B)
186450pdCas12b(RuvC)Encodes catalytically inactive Cas12b (E848A, D977A) from A. acidoterrestris (type V-B)
186451pCas12b(PI)Encodes inactive Cas12b containing mutation in the PI domain (R122A, G143P) from A. acidoterrestris (type V-B)
186452pTargetused as source for acquisition of new spacers
186454pTarget_PS_VApTarget containing protospacer for FnCas12a targeting
186455pTarget_priming_VApTarget containing protospacer with a mismatch on position 1, for FnCas12a targeting
186456pTarget_PS_VBpTarget containing protospacer for AaCas12b targeting

Antibodies from Article