MSMEG0661 (MSMEG0661)

background  +5   0  -5 -10 -15 sgRNA L2FC (+ATc/–ATc) TnSeq Gene Essentiality Predictions (1, 2) Essential Growth Defect Non-Essential Growth Advantage Unknown Operon Legend (3) Operon (1) PMID: 28096490 (2) PMID: 31388080 (3) PMID: 26536359 MSMEG0658 MSMEG0659 MSMEG0660 MSMEG0661 MSMEG0662 MSMEG0663 MSMEG0664
Species: M. smegmatis
Strain: MC2-155
PATRIC Annotation: -
Synonyms:
M. tuberculosis homologs hemL, argD, rocD2, bioA, gabT
M. abscessus homologs MAB3978c, MAB2339, MAB4417c, MAB2279, MAB3841
Gene Start: 742519
Gene End: 743907
Vulnerability Index (VI): -0.1140
VI Lower Bound: -0.2190
VI Upper Bound: -0.0160

Essentiality:

Call Condition Type Source Notes
Non Essential in vitro TnSeq [Akusobi et al. (2022)]
Non Essential in vitro CRISPRi [Bosch & DeJesus et al. (2021)]


DNA Sequence help
TGGGGGCCGACGACGACGGCGGACCGCCCTTGGAGCACGCGTCGAGCAGCACGCCCAGCGCCGGGGCGGTCGCGGCGATCAACGCGGCACGCGTCAGGAACTGTCTACGGGTGGGCCCGGACCGGGGCATCCCGTGCGGCAGTGCGTTTCCGGTGGGTCCAGGCATACCGGCCTCCTGCGGTCCGCGTCTTGTCGCTGGTTGGACTCTCGCATAGACTGAACGCTCAGTCAAGGTACAAAACCGCACATCATCTGCTTTTTTGTCGGATGCGAGCACGTACCTGAGTGGAGGTAATCCAGTGTCCTCGTCAATCGTCGGCTCCGCAGCGCAAGAGGTCGAGAGCGACCGGGTCGACAGACTGATCGCCGACGAGGAACAGGTGTTCCTGCGCCGTCAACCGCGCAGCGCGGAATTCATCGCCCGCGCCCGTGCGCACCTCGCCGGTGGTGCGACGTCCAACTGGCAGATCGCCCAGCCGCAGGCCGTGTGGATGAGCCACGGGTACGGGTCCAAGGTGTACGACGTCGACGGGACGGAGTACGTCGACATGCACGGCGGCTACGGCGCCGCCATCGCCGGGCACGGCCACCCCGCGATCGTCGCGGCCGTGAGCGAACAGGTGCGTCGTGGAACGCATTTCGCACAACCGACCGAGAACGCGATCTGGATCGCCGAGGAACTCTCGCGGCGCTTCGGACTGCCGCTGTGGCGTTTCGCCAACTCGGGGACCGAGGCGACCATGGACGCGGTCCACCTGGCGCGGGCCGTCACCGGACGCGATCTGATCATCAAGGTCGAGGGCTGCTACCACGGTCATCACGACTCGGTGCAGGTTTCCGTGCTGCCCGAAGCCGACGAGGTCGGTCCCCGCGCACACCCGGTCGGTGTTCCCGGCACCAGCGGTATCCCCGAGGCGATCCGCAACCTCGTCGTCGTGGTCCCGTTCAACGACCCCGAGGCCGTGGCGCGGGCGCTCACCGAGCACCGCGGACAGGTGGCCGCGATGATCCTCGAACCGGTGATGATGAACGCGGGCATCATCCCGCCGGCCGACGGCTACCTCGCCGAGATCCGGGATCTGCTGCACGAGGCCGGATCGCTGCTGATCTACGACGAGGTCAAGACCGGCTTCACCGCCGGCCCGGGCGGCATCACCGCGTTGAGCGGGGTGGTGCCCGACATCGTCTGTCTCGCCAAGGCGCTCGGCGGCGGTATCGCCGTCGCGGCCATCGGCGGCACCACGCAGGTCATGTCCGCCATCGCCGACGGGCGCTACGAGCAGGTCGGCACCTTCAACGGCAACCCGCTGGCGATGGCGGCCACCCGCGCGACGCTGAGCGAGGTGCTCACCGACGAGTCGTACGCGCACCTGAACCGCCTCGCGGGCGACCTGCGCGCGCGGTTGGCGAGCGTGATCGCCGAGCACGGGGTTGACTGGCACGTGGTGTCCGTCGGGGCGAAAGGGTGCGTGACCTTCCGCAAGGAACCGGTGCGGGAATATCGTGATTTCCTGCAGATCGACGACCGCCTCGGCCATCTGCACTGGCTGATGCAGCACAACGGCGGTGTTTTCCTGCCGCCATGGGGGAAGGTGGAGCAATGGCTGCTGTCCGTCCAGCACGACACCGCCGATGTCGACCGGTTCGTCGCCAACTTCGCACGCCTCGCCGCGGCGGTGACCCGCTGA
Vulnerability Assesment help
[Full list of passaging experiments]

Experiment: We designed an M. tuberculosis CRISPRi library (RLC12) to target all annotated Mtb genes with single guide RNAs (sgRNAs) of varying predicted knockdown efficiencies. In parallel, we constructed a M. smegmatis CRISPRi library (RLC11) to target all annotated Msmeg genes, similar to the Mtb sgRNA library. RLC12 was transformed into M. tuberculosis H37Rv; RLC11 into M. smegmatis mc2-155. We passaged both CRISPRi libraries for nearly 30 generations in the presence or absence of the CRISPRi-inducer anhydrotetracycline (ATc). At the indicated timepoints, genomic DNA was harvested and the representation of individual sgRNAs was counted using Illumina sequencing. Here, we show sgRNA log2 fold-change values (L2FC) (plus versus minus ATc) over consecutive generations for sgRNAs targeting the gene you selected. sgRNAs are color-coded based on their predicted strength from blue (strength=0; weak) to red (strength=1; strong).

In order to quantify gene vulnerability, data were subsequently analyzed with a hierarchical model at the sgRNA level (two-line model) and gene level (Hill curve). The vulnerability quantification is shown below. [More information]

M. smegmatis Reference Fitness Experiment M. tuberculosis H37Rv Reference Fitness Experiment M. tuberculosis HN878 Reference Fitness Experiment
 
 
M. smegmatis Gene-Level Logistic Regression M. tuberculosis H37Rv Gene-Level Logistic Regression (Most similar ortholog) M. tuberculosis HN878 Gene-Level Logistic Regression (Most similar ortholog)
 
Vulnerability Index (VI) -0.1140 -6.2490
VI Lower Bound -0.2190 -6.5250
VI Upper Bound -0.0160 -5.9860

Specific References

General References