Neisseria gonorrhoeae (Gc, gonococcus) is a Gram-negative, diplococci bacteria and the primary etiological agent of gonorrhea disease. Gonorrhea is the second most prevalent reportable sexually transmitted infection, and there has yet to be a clinically approved vaccine to prevent its spread. As such, gonorrhea represents a major global public health burden. Thus, we need new tools to better interrogate Gc function to develop more efficacious therapies against this bacterium. CRISPR systems (Clustered Regularly Interspaced Short Palindromic Repeats) are prokaryotic adaptive immune systems that protect bacteria from potentially harmful foreign DNA and are regularly utilized as DNA editing tools. In addition to generating mutants, CRISPR-Cas has also been repurposed as a gene repression platform known as CRISPR-interference (CRISPRi). With CRISPRi, an enzymatically deactivated Cas protein complex is directed by its CRISPR guide RNA to a specific DNA sequence where it can sterically block RNA polymerase binding and transcription. I have established an Isopropyl β-D-1-thiogalactopyranoside (IPTG)-inducible, CRISPRi system derived from the commensal Neisseria lactamica’s Type I-C CRISPR-Cas for use in Gc. Importantly, the Cas3 nuclease is missing, rendering it unable to cleave target DNA, but it still can serve as a locus-specific DNA binding and transcription repression machinery. As a proof in principle for using this CRISPRi system, I targeted the constitutively expressed FA1090 opaD gene as it is highly expressed and generates a distinct opaque colony phenotype. 3 I fine-tuned the knockdown magnitude using different IPTG concentrations, developed an efficient guide-switching method, and used this CRISPRi system to knockdown an entire gene family. I have used CRISPRi to target essential genes to generate conditional lethal strains, demonstrating that we can use CRISPRi to study the function of essential genes that cannot be deleted. This new CRISPRi tool has allowed us to interrogate the function of essential yet understudied prophage genes within the Gc genome. Bacteriophages, or bacteria-infecting viruses, are nature’s most abundant entities. Despite the many anti-phage defense systems encoded by bacteria to prevent phage infection, prophage can significantly contribute to the physiology and pathogenicity of its host. Previous bioinformatic analyses identified nine prophage islands (Ngoɸ1-9) in the FA1090 Gc isolate genome. Ngoɸ1-5 are predicted to encode double-strand DNA (dsDNA) phage, while Ngoɸ6-9 are predicted to encode filamentous phage. Of the dsDNA prophage islands, Ngoɸ1-3 are predicted to be most intact. Two previous saturating transposon-sequencing screens using two Gc isolates predicted three phage-like repressors (cI orthologs) encoded in Ngoɸ1 (ngo0479), Ngoɸ2 (ngo1116), and Ngoɸ3 (ngo1630) as being essential. Here, I show that these three genes are weak paralogs and confirm that these genes are essential as a CRISPRi knockdown of each ortholog is lethal to the bacteria. The repression of each cI ortholog leads to the significant induction of phage gene expression in Ngoɸ1-3. Finally, the knockdown of ngo0479, ngo1116, and ngo1630 leads to the production of Siphoviridae-like phage particles. This work marks an important initial step in studying the interaction between Gc and its resident phage.