# Development of CRISPR/dCas-based epigenetic gene regulation tools in malaria parasite

> **NIH NIH R21** · UNIVERSITY OF SOUTH FLORIDA · 2021 · $186,875

## Abstract

PROJECT SUMMARY
The development of resistance in the human malaria parasite Plasmodium falciparum to
essentially all commonly used antimalarial drugs demands increased efforts in drug discovery. A
better understanding of the parasite’s molecular and cellular pathways will help identify novel
drug targets. Although the parasite’s genome has been sequenced more than a decade ago,
the functions of more than half of the genes remain unknown. A major bottleneck towards
functional studies in P. falciparum is the low genetic recombination efficiency. In this proposal, a
new epigenetic gene engineering platform will be designed by taking the advantage of the
CRISPR/Cas9 technology for targeted gene engineering. By fusing either an epigenetic
activator or silencer to a nuclease-deficient Cas9 enzyme (dCas9) and guiding the recombinant
dCas9 to the transcriptional start site of the gene by specific single guide RNA (sgRNA), the
efficient up- or down-regulations of the targeted genes have been achieved. Optimization of this
system to enhance the robustness and precision of timing is critical to meet the requirements for
studying essential genes in this parasite. First, TetR and Cre/loxP inducible modules and
multiplexed gRNAs will be integrated into this system. Second, a complementary gene
regulation system employing the dCas12a (Cpf1), a new Cas with desired features for genetic
engineering in AT-rich genomes like that of P. falciparum, will be engineered and validated.
Systematic comparison of the performance of these two dCas gene regulation systems using a
suite of selected genes expressed at different development stages and with different
transcriptional levels will provide pivotal guidance on future efficient use of the systems. It is
anticipated that this versatile CRISPR/dCas-based epigenetic gene regulation system would
find broad applications in functional genomic studies in P. falciparum.

## Key facts

- **NIH application ID:** 10084810
- **Project number:** 5R21AI149202-02
- **Recipient organization:** UNIVERSITY OF SOUTH FLORIDA
- **Principal Investigator:** Jun Miao
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $186,875
- **Award type:** 5
- **Project period:** 2020-01-13 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10084810

## Citation

> US National Institutes of Health, RePORTER application 10084810, Development of CRISPR/dCas-based epigenetic gene regulation tools in malaria parasite (5R21AI149202-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10084810. Licensed CC0.

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