Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (7): 62-69.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1592

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Optimization and Application of the Chromatin Immunoprecipitation Based on Rice Protoplast

SHI Jia(), ZHU Xiu-mei, XUE Meng-yu, YU Chao, WEI Yi-ming, YANG Feng-huan, CHEN Hua-min()   

  1. State Key Laboratory for Biology of Plant Disease and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Science,Beijing 100193
  • Received:2021-12-25 Online:2022-07-26 Published:2022-08-09
  • Contact: CHEN Hua-min E-mail:qwer1768004860@163.com;chenhuamin@caas.cn

Abstract:

Transcription factors(TFs)are important regulatory proteins that control gene expression and regulate various biological functions in plant. Chromatin immunoprecipitation(ChIP)is a very widely used technique to investigate the direct target genes of TFs. However,ChIP is always limited by antibody specificity or time-consuming construction of genetic materials. In this paper,the chromatin immunoprecipitation system based on rice protoplasts(ChIP-RP)was established by optimizing the transient transformation of rice protoplasts,formaldehyde fixation and ultrasonic fragmentation of immunoprecipitated nucleic acid. Moreover,the enrichment of target genes by OsNF-YA4 protein,a transcription factor of rice,were verified via ChIP-RP method. This technical system will greatly reduce the limitations of antibody specificity or constructing stable genetic materials,facilitate rapid screening and verification of direct target genes of rice transcription factors,and promote the deciphering of the molecular mechanism of rice transcription factor regulatory functions.

Key words: rice protoplast, chromatin immunoprecipitation, transcription factor, OsNF-YA4, ChIP-RP