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Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage.
Kulcsar PI, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N, Fodor E, Welker E
Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.
PubMed

Plasmids from Article

ID Plasmid Purpose  
92102pX330-Flag-SpCas9-HF1 (without sgRNA)Expression plasmid for human codon-optimized high-fidelity SpCas9-HF1, px330-like backbone (without U6-sgRNA coding sequence)
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92110HeFm1SpCas9Expression plasmid for human codon-optimized increased fidelity HeFm1SpCas9 (without U6-sgRNA coding sequence)
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92111HeFm2SpCas9Expression plasmid for human codon-optimized increased fidelity HeFm2SpCas9 (without U6-sgRNA coding sequence)
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92112pX330-HA-dSpCas9Expression plasmid for human codon-optimized dead/inactive SpCas9 (without U6-sgRNA coding sequence)
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92113pX330-Flag-dSpCas9Expression plasmid for human codon-optimized dead/inactive SpCas9 (without U6-sgRNA coding sequence)
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92114pX330-Flag-deSpCas9Expression plasmid for human codon-optimized dead/inactive increased fidelity eSpCas9 (1.1) (without U6-sgRNA coding sequence)
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92115pX330-Flag-dSpCas9-HF1Expression plasmid for human codon-optimized dead/inactive high-fidelity SpCas9-HF1 (without U6-sgRNA coding sequence)
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92116pX330-Flag-dHeFSpCas9Expression plasmid for human codon-optimized dead/inactive increased fidelity HeFSpCas9 (without U6-sgRNA coding sequence)
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92117pET-deSpCas9-VP64-6xHisExpression of dead/inactive increased fidelity eSpCas9 (1.1)-VP64-6xHis in bacterial cells
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92118pET-dSpCas9-HF1-VP64-6xHisExpression of dead/inactive increased fidelity SpCas9-HF1-VP64-6xHis in bacterial cells
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92119pET-dHeFSpCas9-VP64-6xHisExpression of dead/inactive increased fidelity HeFSpCas9-VP64-6xHis in bacterial cells
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92120U6-sgRNA(MS2)_EF1a-MS2-P65-HSF1Expression plasmid for both MS2-P65-HSF1 activator helper complex and sgRNA with two MS2 loops at tetraloop and stemloop 2 contains BsaI sites for insertion of spacer sequences.
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92353pX330-Flag-wtSpCas9 (without sgRNA)Expression plasmid for human codon-optimized wild-type SpCas9 (without U6-sgRNA coding sequence)
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92354pX330-Flag-eSpCas9 (without sgRNA)Expression plasmid for human codon-optimized high-fidelity eSpCas9 (without U6-sgRNA coding sequence)
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92355HeFSpCas9Expression plasmid for human codon-optimized increased fidelity HeFSpCas9 (without U6-sgRNA coding sequence)
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