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Clustered regularly interspaced palindromic repeats (CRISPR) and their associatedcas genes have been demonstrated to regulate self-genes and virulence in many pathogens. In this study, we found that inactivation ofcas9 caused reduced adhesion and intracellular survival of the piscineStreptococcus agalactiae strain GD201008-001 and significantly decreased the virulence of this strain in zebrafish and mice. Further investigation indicated that theregR transcriptional regulator was upregulated in the Δcas9 mutant. AsregR mediates the repression of hyaluronidase, a critical factor involved in opening the blood-brain barrier (BBB) in mice,cas9 -mediated repression ofregR transcription is important forS. agalactiae to open the BBB and thereby cause meningitis in animals. This study expands our understanding of endogenous gene regulation mediated by CRISPR-Cas systems in bacteria.

cas9 Enhances Bacterial Virulence by Repressing the regR Transcriptional Regulator in Streptococcus agalactiae