10xhis mbp  (Addgene inc)

Journal: Antimicrobial Agents and Chemotherapy

Article Title: EfpA is required for regrowth of Mycobacterium tuberculosis following isoniazid exposure

doi: 10.1128/aac.00261-24

Figure Lengend Snippet: Strains and plasmids used

Article Snippet: pRH2502 , Integrative plasmid originating from the pTC-0X-1L backbone, expressing S. pyogenes dCas9 under control of the tetracyline-inducible tetRO promoter (uv15 tetO ) , ( ); pRH2502 (Addgene plasmids # 84379) .

Techniques: Plasmid Preparation, Transformation Assay, Control, Sequencing, Knockdown, Expressing, Clone Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: EfpA is required for regrowth of Mycobacterium tuberculosis following isoniazid exposure

doi: 10.1128/aac.00261-24

Figure Lengend Snippet: A two-plasmid dCas9–sgRNA CRISPRi system adapted to repress efpA expression. Schematic of the single-guide RNA chimeric structure consisting of the terminator region originating from Streptococcus pyogenes, the dCas9 handle region (to enable binding to the dCas9 enzyme), and the sequence complementary to the efpA target sequence at the 5’ end of the gene. The protospacer adjacent motif (PAM) sequence (CCN) is adjacent to the 5’ end of the target sequence ( A ). Schematic of the CRISPRi genes on two plasmids coding for the CRISPRi system: pRH2502, which encodes the dCas9 endonuclease enzyme under the control of a uv tetO promoter, and pRH2521- efpA , which encodes the efpA sgRNA under the control of a p myc1tetO promoter ( B ).

Article Snippet: pRH2502 , Integrative plasmid originating from the pTC-0X-1L backbone, expressing S. pyogenes dCas9 under control of the tetracyline-inducible tetRO promoter (uv15 tetO ) , ( ); pRH2502 (Addgene plasmids # 84379) .

Techniques: Plasmid Preparation, Expressing, Binding Assay, Sequencing, Control