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Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants.
Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26. PubMed Article

Plasmids from Article

ID Plasmid Purpose
139987pCMV-T7-SpCas9-P2A-EGFP (RTW3027)CMV and T7 promoter expression plasmid for human codon optimized SpCas9 with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139988pCMV-T7-SpG-P2A-EGFP (RTW4177)CMV and T7 promoter expression plasmid for human codon optimized SpG(D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139989pCMV-T7-SpRY-P2A-EGFP (RTW4830)CMV and T7 promoter expression plasmid for human codon optimized SpRY(A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139990pCMV-T7-SpCas9-VQR-P2A-EGFP (RTW3520)CMV and T7 promoter expression plasmid for human codon optimized SpCas9-VQR(D1135V/R1335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139991pCMV-T7-SpCas9-VRER-P2A-EGFP (RTW3160)CMV and T7 promoter expression plasmid for human codon optimized SpCas9-VRER(D1135V/G1218R/R1335E/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139992pCMV-T7-SpCas9-VRQR-P2A-EGFP (RTW3161)CMV and T7 promoter expression plasmid for human codon optimized SpCas9-VRQR(D1135V/G1218R/R1335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139993pCMV-T7-xCas9(3.7)-P2A-EGFP (RTW3576)CMV and T7 promoter expression plasmid for human codon optimized xCas9(3.7)(A262T/R324L/S409I/E480K/E543D/M694I/E1219V) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139994pCMV-T7-SpCas9-NG-P2A-EGFP (RTW3677)CMV and T7 promoter expression plasmid for human codon optimized SpCas9-NG(L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139995pCMV-T7-SpCas9-HF1-P2A-EGFP (RTW3505)CMV and T7 promoter expression plasmid for human codon optimized SpCas9-HF1(N497A/R661A/Q695A/Q926A) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139996pCMV-T7-SpG-HF1-P2A-EGFP (RTW5000)CMV and T7 promoter expression plasmid for human codon optimized SpG-HF1(SpG=D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R;HF1=N497A/R661A/Q695A/Q926A) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139997pCMV-T7-SpRY-HF1-P2A-EGFP (RTW5008)CMV and T7 promoter expression plasmid for human codon optimized SpRY-HF1(SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R;HF1=N497A/R661A/Q695A/Q926A) with a c-terminal bi-partite NLS, 3x flag tag, and P2A-EGFP
139998pCAG-CBE4max-SpG-P2A-EGFP (RTW4552)CAG promoter expression plasmid for human codon optimized BE4max C-to-T base editor with SpG(D10A/D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R) and P2A-EGFP
139999pCAG-CBE4max-SpRY-P2A-EGFP (RTW5133)CAG promoter expression plasmid for human codon optimized BE4max C-to-T base editor with SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R) and P2A-EGFP
140000pCAG-CBE4max-xCas9(3.7)-P2A-EGFP (RTW4637)CAG promoter expression plasmid for human codon optimized BE4max C-to-T base editor with xCas9(3.7)(D10A/A262T/R324L/S409I/E480K/E543D/M694I/E1219V) and P2A-EGFP
140001pCAG-CBE4max-SpCas9-NG-P2A-EGFP (RTW4554)CAG promoter expression plasmid for human codon optimized BE4max C-to-T base editor with SpCas9-NG(D10A/L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) and P2A-EGFP
140002pCMV-T7-ABEmax(7.10)-SpG-P2A-EGFP (RTW4562)CMV and T7 promoter expression plasmid for human codon optimized ABEmax(7.10) A-to-G base editor with SpG(D10A/D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R) and P2A-EGFP
140003pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)CMV and T7 promoter expression plasmid for human codon optimized ABEmax(7.10) A-to-G base editor with SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R) and P2A-EGFP
140004pCMV-T7-ABEmax(7.10)-xCas9(3.7)-P2A-EGFP (RTW4644)CMV and T7 promoter expression plasmid for human codon optimized ABEmax(7.10) A-to-G base editor with xCas9(3.7)(D10A/A262T/R324L/S409I/E480K/E543D/M694I/E1219V) and P2A-EGFP
140005pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564)CMV and T7 promoter expression plasmid for human codon optimized ABEmax(7.10) A-to-G base editor with SpCas9-NG(D10A/L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) and P2A-EGFP

Antibodies from Article