The ribosomal RNA promoter is more efficient than the U6 promoter to drive gRNA expression in Leishmania. The ribosomal RNA promoter is more efficient.

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The ribosomal RNA promoter is more efficient than the U6 promoter to drive gRNA expression in Leishmania. The ribosomal RNA promoter is more efficient than the U6 promoter to drive gRNA expression in Leishmania. (A) Different LdMT gRNAa expression vectors using the L. donovani rRNA promoter (LdrRNAP), L. donovani U6 promoter (LdU6), and human U6 promoter (human U6), respectively. Unlike LdrRNAP, which initiates transcription at the T residue site, L. donovani and human U6 promoters use a G residue to initiate transcription. As the U6 promoter terminates transcription at the 3′ end, a poly(T) (5-6) site of the gRNA coding sequence, HDV ribozyme (H) in this LdU6 promoter vector is not required, though it is able to process any of the passthrough transcripts. (B) The MLF resistance rates of L. donovani cells transfected with the various promoter-driving gRNAa expression vectors. The miltefosine (MLF) resistance rates were determined by limiting dilution as detailed in reference 7. Wen-Wei Zhang et al. mSphere 2017; doi:10.1128/mSphere.00340-16