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Engineering domain-inlaid SaCas9 adenine base editors with reduced RNA off-targets and increased on-target DNA editing.
Nguyen Tran MT, Mohd Khalid MKN, Wang Q, Walker JKR, Lidgerwood GE, Dilworth KL, Lisowski L, Pebay A, Hewitt AW
Nat Commun. 2020 Sep 25;11(1):4871. doi: 10.1038/s41467-020-18715-y.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
1353401010-AIDx-CPExpression of circular permutant of nSpyCas9 at amino acid position 1010 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus.
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1353411029-AIDx-CPExpression of circular permutant nSpyCas9 at amino acid position 1029 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus.
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1353421058-AIDx-CPExpression of circular permutant of nSpyCas9 at amino acid position 1058 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus.
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135343SpyCas9-AIDxExpression of wildtype nSpyCas9 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus.
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135344AIDx-ID-202Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 202 of nSpyCas9 used for base editing
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135345AIDx-ID-208Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 208 of nSpyCas9
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135346AIDx-ID-468Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 468 of nSpyCas9
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135347AIDx-ID-1058Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 1058 of nSpyCas9 with uracil DNA glycosylase inhibitor (UGI) at the C terminus
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135348BE3-NLS-NLS-P2A-BLASTExpression of BE3 (Base editor generation three) with dual C-terminal NLS in a single operon encoding P2A blasticidine resistance
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135349S2 1058 AIDx no UGIExpression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 1058 of nSpyCas9 lacking uracil DNA glycosylase inhibitor.
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135350S2 1058 APOBEC3 no UGIExpression of intradomain insertion of rat cytosine deaminase, rAPOBEC3, at position 1058 of nSpyCas9 lacking uracil DNA glycosylase inhibitor.
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135352S1 1029 CP APOBEC3 no UGIExpression of circular permutant of nSpyCas9 lacking uracil DNA glycosylase inhibitor at amino acid position 1029 encoding rat cytosine deaminase, rAPOBEC3 at the N-terminus
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135354S1 1029 AIDx with UGIExpression of nSpyCas9 encoding truncated human cytosine deaminase hAIDx at amino acid 1029 and uracil DNA glycosylase inhibitor (UGI) at the C terminus
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135355pCMV-ABEMAX-NLS-P2A_BLASTExpression of ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) with dual C-terminal NLS in a single operon encoding P2A blasticidine resistance
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135356S2 1058 ABEMaxExpression of intradomain insertion of ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at position 1058 of nSpyCas9
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135357S1 1029 CP ABEMax strategy 1Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at the N-terminus
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135358S1 1029 CP ABEMax strategy 2Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding de-coupled ABEMax (wildtype Tada monomer at N terminus, evolved Tada monomer at C terminus)
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135359S1 1029 CP ABEMax strategy 3Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at the C-terminus
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135360S1 1029 miniABEMax V82G strategy1Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding miniABEMax (evolved Tada monomer with V82G) at the N-terminus
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135361S2 1058 V82G ABEMaxExpression of intradomain insertion of miniABEMax (evolved Tada monomer with V82G) at position 1058 of nSpyCas9
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135362miniABEMax V82G wtExpression of nSpyCas9 with N-terminal miniABEMax (V82G)
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135363S1 1029 V82G ABEMax Strategy 3Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding miniABEMax (evolved Tada monomer with V82G) at the N-terminus
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135364ABEMax nSaCas9Expresion of SaCas9 nickase (D10A) with ABEMax at the N terminus
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135365miniABEMax V82G nSaCas9Expression of SaCas9 nickase (D10A) with single evolved TadA monomer (ABEMax) with a V82G substitution
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135366nSaCas9 D127 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (D127 )
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135367nSaCas9 D126 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (D126 )
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135368nSaCas9 E739 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E739 )
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135369nSaCas9 E743 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E743 )
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135370nSaCas9 E745 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E745 )
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135371nSaCas9 F733 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (F733 )
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135372nSaCas9 N730 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N730 )
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135373nSaCas9 E125 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E125 )
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135374nSaCas9 E131 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E131 )
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135375nSaCas9 L132 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (L132 )
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135376nSaCas9 N130 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N130 )
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135377nSaCas9 E735 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E735 )
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135379nSaCas9 I744 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (I744 )
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135380nSaCas9 G129 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (G129 )
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135381nSaCas9 H119 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (H119 )
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135382nSaCas9 M732 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (M732 )
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135383nSaCas9 M741 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (M741 )
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135384nSaCas9 N120 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N120 )
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135385nSaCas9 P742 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (P742 )
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135386nSaCas9 Q731 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (Q731 )
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135387nSaCas9 Q737 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (Q737 )
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135388nSaCas9 S740 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (S740 )
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135389nSaCas9 T128 miniABEMax V82GExpression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (T128 )
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