Project description:WT M. abscessus ATCC_19977 was used to construct a targeted deletion mutant lacking the sRNA B11 and a complemented version of the mutant with ectopic expression of B11 from its native promoter at the L5 site. The deletion mutant was also complemented with versions of B11 that lacked a single C from the first loop or had an extra C in the second loop. All strains were grown to log phase and the extracted RNA was used to perform expression profiling by RNAseq.
Project description:Understanding the regulatory roles of small RNAs (sRNAs) in Mycobacterium marinum is crucial for elucidating its pathogenesis. Here, we present transcriptome profiles of M. marinum strains with deletions and completions of sRNA B11. Through RNA sequencing analysis, we identified significant alterations in gene expression patterns between the B11-deleted and completed strains.
Project description:To study the role of NRAS mutations in cell proliferation and self-renewal in acute myeloid leukemia (AML), the human AML cell line, THP1, was modified to replace its naturally occurring heterozygous NRAS-G12D mutation with a doxycycline(dox)-inducible heterozygous NRAS-G12V mutation. The endogenous copies of the NRAS-G12D allele were deleted using CRISPR/Cas9 after a dox-inducible, CRISPR resistant, NRAS-G12V transgene was introduced into the THP1 cell line. The resulting cell line was named B11. RNA-seq data confirmed that endogenous NRAS G12D was successfully replaced by dox-inducible exogenous NRAS G12V in the B11 cell line. As expected, depletion of dox induced G1 cell cycle arrest. Interestingly, the B11 cells experienced ten-times higher expression of NRAS induced G2/S-phase cell cycle arrest. Forty-nine genes were identified as signaling responsible genes associated with high expression of NRAS.
