Genome-wide Identification of Immune Regulatory Networks: Comparative Performance of RNAi and CRISPRi in Host Factor Discovery
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Viruses systematically exploit numerous host immune regulatory factors to establish infections, yet endogenous suppression mechanisms remain inadequately characterized. This review examines RNA interference (RNAi) and CRISPR interference (CRISPRi) screening methodologies for systematically identifying host factors that negatively regulate immune signaling pathways, addressing fundamental questions regarding the molecular mechanisms of immune suppression through ubiquitin regulation, phosphorylation control, and transcriptional repression, Secondly, the discovery of non-canonical regulatory proteins through unbiased functional genomics. This systematic review employed PRISMA-guided literature analysis of peer-reviewed publications from PubMed, Embase, Web of Science, and Cochrane databases (inception-2024), utilizing structured Boolean search strategies with immune-specific MeSH terms, followed by quantitative meta-analysis of screening efficiency data and qualitative synthesis of mechanistic findings through thematic coding and comparative framework analysis. Rigorous comparative analysis demonstrates CRISPRi's better performance with enhanced knockdown efficiency (85% ± 12%) versus RNAi's (60% ± 25%), reduced off-target effects (0.3-0.8% versus 15-30%), and improved temporal stability via direct transcriptional interference. Comprehensive screening reveals diverse immunosuppressants including ubiquitin-editing enzymes (A20, CYLD), transcriptional repressors (BCL6), and metabolic mediators (CH25H), with robust cross-platform validation. Clinical evidence demonstrates that immune suppressor dysregulation correlates with altered viral susceptibility patterns, validating therapeutic potential. This systematic mapping of regulatory networks through advanced functional genomics enables development of innovative host-directed interventions for infectious diseases and cancer immunotherapy applications.
