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Vol. 14, Issue 1 (2025)

CRISPRi: Based identification of tuberculosis drug targets and microcrystalline cellulose-enhanced drug delivery for sustained therapeutic efficacy

Author(s):
Elena Fernández Torres
Abstract:
Tuberculosis (TB) remains a major global health challenge, exacerbated by the rise of multidrug-resistant Mycobacterium tuberculosis (Mtb). The inefficacy of existing drugs due to resistance mechanisms necessitates novel drug targets and optimized drug delivery systems. This study aimed to identify essential drug targets in Mtb using CRISPR interference (CRISPRi) screening and to evaluate the efficacy of microcrystalline cellulose (MCC)-based formulations for sustained drug delivery. A genome-wide CRISPRi library was constructed using dCas9-mediated transcriptional repression, and gene knockdown efficiency was assessed using qPCR and RNA sequencing (RNA-Seq). The impact of gene suppression on bacterial survival and drug susceptibility was evaluated using broth dilution assays and colony-forming unit (CFU) enumeration. MCC-based isoniazid formulations were developed using a wet granulation method and characterized via scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). In vitro drug release profiles were assessed using a USP Dissolution Apparatus II, and statistical analysis, including ANOVA and Pearson correlation, was performed to determine significant trends.
The results showed that high CRISPRi knockdown efficiency correlated with reduced bacterial survival (r = -0.78, p< 0.0001), indicating that essential genes were successfully identified. A positive correlation (r = 0.61) between bacterial survival and rifampicin MIC values confirmed that knockdowns affected drug susceptibility. MCC-based formulations demonstrated sustained drug release over 24 hours, with a strong negative correlation (-0.68) between MCC drug release and bacterial survival, confirming prolonged antimicrobial activity. The study concludes that CRISPRi is a powerful tool for tuberculosis drug target identification, while MCC-based formulations offer a promising strategy for sustained drug delivery. Future research should integrate in vivo pharmacokinetics, whole-genome sequencing, and advanced drug carriers to further optimize tuberculosis treatment strategies.
Pages: 94-98  |  47 Views  29 Downloads


The Pharma Innovation Journal
How to cite this article:
Elena Fernández Torres. CRISPRi: Based identification of tuberculosis drug targets and microcrystalline cellulose-enhanced drug delivery for sustained therapeutic efficacy. Pharma Innovation 2025;14(1):94-98.

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