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Heterologous Expression and Purification of CRISPR-Cas12a/Cpf1

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BIO-PROTOCOL
卷 8, 期 9, 页码 -

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BIO-PROTOCOL
DOI: 10.21769/BioProtoc.2842

关键词

CRISPR-Cas; Cas12a; Cpf1; Protein purification

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资金

  1. Netherlands Organization of Scientific Research (NWO) [714.015.001]
  2. Swiss National Science Foundation (SNSF) [SNSF 31003A_149393]
  3. European Molecular Biology Organization (EMBO) [EMBO ALTF 179-2015, EMBO aALTF 509-2017]

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This protocol provides step by step instructions (Figure 1) for heterologous expression of Francisella novicida Cas12a ( previously known as Cpf1) in Escherichia coli. It additionally includes a protocol for high-purity purification and briefly describes how activity assays can be performed. These protocols can also be used for purification of other Cas12a homologs and the purified proteins can be used for subsequent genome editing experiments. [GRAPHICS]

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