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.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 135340 | 1010-AIDx-CP | Expression of circular permutant of nSpyCas9 at amino acid position 1010 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus. |
| 135341 | 1029-AIDx-CP | Expression of circular permutant nSpyCas9 at amino acid position 1029 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus. |
| 135342 | 1058-AIDx-CP | Expression of circular permutant of nSpyCas9 at amino acid position 1058 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus. |
| 135343 | SpyCas9-AIDx | Expression of wildtype nSpyCas9 encoding truncated human cytosine deaminase, hAIDx, at the C-terminus. |
| 135344 | AIDx-ID-202 | Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 202 of nSpyCas9 used for base editing |
| 135345 | AIDx-ID-208 | Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 208 of nSpyCas9 |
| 135346 | AIDx-ID-468 | Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 468 of nSpyCas9 |
| 135347 | AIDx-ID-1058 | Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 1058 of nSpyCas9 with uracil DNA glycosylase inhibitor (UGI) at the C terminus |
| 135348 | BE3-NLS-NLS-P2A-BLAST | Expression of BE3 (Base editor generation three) with dual C-terminal NLS in a single operon encoding P2A blasticidine resistance |
| 135349 | S2 1058 AIDx no UGI | Expression of intradomain insertion of truncated human cytosine deaminase, hAIDx, at position 1058 of nSpyCas9 lacking uracil DNA glycosylase inhibitor. |
| 135350 | S2 1058 APOBEC3 no UGI | Expression of intradomain insertion of rat cytosine deaminase, rAPOBEC3, at position 1058 of nSpyCas9 lacking uracil DNA glycosylase inhibitor. |
| 135352 | S1 1029 CP APOBEC3 no UGI | Expression of circular permutant of nSpyCas9 lacking uracil DNA glycosylase inhibitor at amino acid position 1029 encoding rat cytosine deaminase, rAPOBEC3 at the N-terminus |
| 135354 | S1 1029 AIDx with UGI | Expression of nSpyCas9 encoding truncated human cytosine deaminase hAIDx at amino acid 1029 and uracil DNA glycosylase inhibitor (UGI) at the C terminus |
| 135355 | pCMV-ABEMAX-NLS-P2A_BLAST | Expression of ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) with dual C-terminal NLS in a single operon encoding P2A blasticidine resistance |
| 135356 | S2 1058 ABEMax | Expression of intradomain insertion of ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at position 1058 of nSpyCas9 |
| 135357 | S1 1029 CP ABEMax strategy 1 | Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at the N-terminus |
| 135358 | S1 1029 CP ABEMax strategy 2 | Expression 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) |
| 135359 | S1 1029 CP ABEMax strategy 3 | Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding ABEMax (wildtype Tada monomer coupled with evolved Tada monomer) at the C-terminus |
| 135360 | S1 1029 miniABEMax V82G strategy1 | Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding miniABEMax (evolved Tada monomer with V82G) at the N-terminus |
| 135361 | S2 1058 V82G ABEMax | Expression of intradomain insertion of miniABEMax (evolved Tada monomer with V82G) at position 1058 of nSpyCas9 |
| 135362 | miniABEMax V82G wt | Expression of nSpyCas9 with N-terminal miniABEMax (V82G) |
| 135363 | S1 1029 V82G ABEMax Strategy 3 | Expression of circular permutant of nSpyCas9 at amino acid position 1029 encoding miniABEMax (evolved Tada monomer with V82G) at the N-terminus |
| 135364 | ABEMax nSaCas9 | Expresion of SaCas9 nickase (D10A) with ABEMax at the N terminus |
| 135365 | miniABEMax V82G nSaCas9 | Expression of SaCas9 nickase (D10A) with single evolved TadA monomer (ABEMax) with a V82G substitution |
| 135366 | nSaCas9 D127 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (D127 ) |
| 135367 | nSaCas9 D126 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (D126 ) |
| 135368 | nSaCas9 E739 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E739 ) |
| 135369 | nSaCas9 E743 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E743 ) |
| 135370 | nSaCas9 E745 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E745 ) |
| 135371 | nSaCas9 F733 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (F733 ) |
| 135372 | nSaCas9 N730 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N730 ) |
| 135373 | nSaCas9 E125 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E125 ) |
| 135374 | nSaCas9 E131 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E131 ) |
| 135375 | nSaCas9 L132 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (L132 ) |
| 135376 | nSaCas9 N130 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N130 ) |
| 135377 | nSaCas9 E735 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (E735 ) |
| 135379 | nSaCas9 I744 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (I744 ) |
| 135380 | nSaCas9 G129 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (G129 ) |
| 135381 | nSaCas9 H119 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (H119 ) |
| 135382 | nSaCas9 M732 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (M732 ) |
| 135383 | nSaCas9 M741 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (M741 ) |
| 135384 | nSaCas9 N120 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (N120 ) |
| 135385 | nSaCas9 P742 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (P742 ) |
| 135386 | nSaCas9 Q731 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (Q731 ) |
| 135387 | nSaCas9 Q737 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (Q737 ) |
| 135388 | nSaCas9 S740 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (S740 ) |
| 135389 | nSaCas9 T128 miniABEMax V82G | Expression of nSaCas9 intradomain insertion of single evolved TadA monomer (V82G) at the specified position (T128 ) |