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Published on: March 2, 2013
Epigenetic editing by CRISPR/dCas9 in Plasmodium falciparum
Bo Xiao1, Shigang Yin1, Yang Hu1
1Unit of Human Parasite Molecular and Cell Biology, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai 200031, P.R. China.
Abstract:
Genetic manipulation remains a major obstacle for understanding the functional genomics of the deadliest malaria parasite Plasmodium falciparum Although the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) system has been successfully applied to introduce permanent changes in the parasite genome, its use is still limited. Here we show that fusing different epigenetic effector domains to a Cas9 null mutant efficiently and specifically reprograms the expression of target genes in P. falciparum By precisely writing and erasing histone acetylation at the transcription start site regions of the invasion-related genes reticulocyte binding protein homolog 4 (rh4) and erythrocyte binding protein 175 (eba-175), respectively, we achieved significant activation of rh4 and repression of eba-175, leading to the switch of the parasite invasion pathways into human erythrocytes. By using the epigenetic knockdown system, we have also characterized the effects of PfSET1, previously identified as an essential gene, on expression of mainly trophozoite- and schizont-specific genes, and therefore regulation of the growth of the mature forms of P. falciparum This epigenetic CRISPR/dCas9 system provides a powerful approach for regulating gene expression at the transcriptional level in P. falciparum.
Insights
Scientists developed a new CRISPR/dCas9 epigenetic system to precisely control gene expression in the malaria parasite Plasmodium falciparum. This breakthrough enables targeted gene activation or repression, advancing functional genomics research for this deadly pathogen.
Area of Science:
- Malaria research
- Parasitology
- Molecular biology
Background:
- Understanding the functional genomics of Plasmodium falciparum, the deadliest malaria parasite, is hindered by genetic manipulation challenges.
- The CRISPR/Cas9 system has limitations for permanent genome modifications in P. falciparum.
Purpose of the Study:
- To develop an efficient and specific method for reprogramming gene expression in P. falciparum using an epigenetic CRISPR/dCas9 system.
- To investigate the role of epigenetic modifications in regulating invasion pathways and parasite growth.
Main Methods:
- Fusing epigenetic effector domains to a Cas9 null mutant.
- Targeting histone acetylation at transcription start sites of invasion-related genes (rh4 and eba-175).
- Utilizing the epigenetic knockdown system to characterize PfSET1 function.
Main Results:
- Successfully activated rh4 and repressed eba-175 expression by precise histone acetylation reprogramming.
- Demonstrated a switch in parasite invasion pathways into human erythrocytes.
- Characterized the role of PfSET1 in regulating trophozoite- and schizont-specific gene expression and parasite growth.
Conclusions:
- The epigenetic CRISPR/dCas9 system offers a powerful approach for transcriptional gene regulation in P. falciparum.
- This system facilitates functional genomics studies and the characterization of essential genes in malaria parasites.
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