Transcriptome analysis of Haloferax volcanii mutant strain Cas8M545A vs wildtype H119

We identified a small subunit protein (Cas11b) of the Cascade Complex in the CRISP-Cas Type I-b System of Haloferax volcanii, which is internally translated from Cas8. We identified a small subunit protein (Cas11b) of the Cascade Complex in the CRISP-Cas Type I-b System of Haloferax volcanii, which is internally translated from Cas8. We aim to investigate the transcripts that are differentially expressed upon mutation of the translation start site of Cas11(Cas8M545A), which leads to loss of Cas11 proteins, to elucidate the cellular functions of this small subunit beyond the essential function for CRISPR-Cas interference that we have experimentally demonstrated. Overall design: We isolated total RNA from Haloferax volcanii Cas8M545A and the wildtype strain H119 at early exponential growth and subjected it to rRNA depletion and then next generation RNA sequencing (RNA-seq) (each in quintuplicate).