Heterologous Expression and Purification of a CRISPR-Cas9-Based Adenine Base Editor

Jan 2, 2023Methods in molecular biology (Clifton, N.J.)

Producing and Purifying a CRISPR-Cas9 Tool That Changes DNA Bases

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Abstract

CRISPR-Cas9-guided adenine base editors (ABEs) can convert A-T base pairs to G-C base pairs in genomic DNA.

  • The delivery of preassembled ABE proteins with guide RNAs may significantly reduce off-target effects compared to other methods.
  • High purity and quantity of ABE-gRNA complexes are essential for effective gene editing using the ribonucleoprotein delivery method.
  • The procedure for expressing and purifying ABEmax, a high-efficiency version of ABE, is outlined.

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