Jingchang Liang

All Publications

• Production of Homogeneous, Functional Zinc-Finger Arrays in High Yield With Two Chromatographic Steps. Bio-protocol Liang, J., Azubel, M., Wang, G., Nie, Y., Kornberg, R. D., Beel, A. J., Matteï, P. J. 2025; 15 (16): e5420

  Abstract

  Zinc-finger (ZF) arrays are compact, sequence-specific polynucleotide-binding domains, which have been used to target the delivery of diverse effector domains, enabling applications such as gene identification, localization, regulation, and editing. To facilitate in vitro applications of ZF arrays, we have developed a general method for their expression and purification. Here, we describe a protocol involving two chromatographic steps that yields homogeneous and functional ZF arrays in milligram quantities. Key features • A general method for expressing and purifying C2H2 ZF arrays in E. coli, compatible with both natural and artificial ZFs. • The His-SUMO tag improves ZF-array solubility, eliminating the need for denaturation and refolding steps. • Simple, two-step purification yields milligram-scale ZF arrays suitable for downstream applications.

  View details for DOI 10.21769/BioProtoc.5420

  View details for PubMedID 40873471

  View details for PubMedCentralID PMC12378415

• A universal method for the purification of C2H2 zinc finger arrays. PloS one Liang, J., Azubel, M., Wang, G., Nie, Y., Kornberg, R. D., Beel, A. J., Mattei, P. J. 2025; 20 (2): e0318295

  Abstract

  Zinc fingers (ZFs) are compact, modular, sequence-specific polynucleotide-binding domains uniquely suited for use as DNA probes and for the targeted delivery of effector domains for purposes such as gene regulation and editing. Despite recent advances in both the design and application of ZF-containing proteins, there is still a lack of a general method for their expression and purification. Here we describe a simple method, involving two chromatographic steps, for the production of homogeneous, functional ZF proteins in high yield (one milligram per liter of bacterial culture), and we demonstrate the generality of this method by applying it to a diverse set of eight C2H2-type ZF proteins. By incorporating a surface-exposed terminal cysteine residue that enables site-specific conjugation with maleimide-activated fluorophores, we confirm the suitability of these probes for in situ labeling of specific DNA sequences in human cells.

  View details for DOI 10.1371/journal.pone.0318295

  View details for PubMedID 39903729

  View details for PubMedCentralID PMC11793764