The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is the latest RNA-guided, endonuclease tool in genome editing which allows for very specific genomic disruption and replacement. The Cas9 Null Mutant NLS Protein (also referred to as Double Mutant) is created by mutating both cleavage domains of the wild type Cas9 (D10A and H840A) and adding a SV40 T antigen nuclear localization sequence (NLS) on the C-terminus of the protein. Such a Cas9 protein retains its ability to bind to genomic DNA through guide RNA (gRNA):genomic DNA base pairing. However, unlike Cas9 Nuclease and Cas9 Nickase, where permanent gene disruption can be achieved, the Cas9 Null Mutant does not introduce any genome modifications. Therefore, this protein can provide a useful negative control for CRISPR experiments. By fusing the Cas9 null mutant with other effector proteins, the CRISPR Cas9 system can expand its role to gene regulation, genome imaging, chromatin or DNA modifications, and chromatin immunoprecipitation. In addition, binding of the Null Mutant can act as a roadblock to hinder transcription, thus offering a useful tool to achieve reversible knock-down of gene expression.