# MSMEG_0918 is not Essential for the Growth of Mycobacterium smegmatis

## TL;DR

This study shows that the gene MSMEG_0918 is not essential for the growth of Mycobacterium smegmatis under standard conditions.

## Contribution

The novelty is demonstrating that MSMEG_0918, unlike its homolog in M. tuberculosis, is not essential for M. smegmatis growth.

## Key findings

- MSMEG_0918 knockdown strains showed no growth differences compared to wild-type M. smegmatis.
- The gene MSMEG_0918 lacks a copper-binding motif and does not affect aerobic growth.
- Generated knockdown strains can be used for further functional studies of MSMEG_0918.

## Abstract

Copper homeostasis plays a crucial role in mycobacteria. In
Mycobacterium tuberculosis
, Rv0474 is a copper-responsive regulator with a copper-binding motif but its homolog in
Mycobacterium smegmatis
, MSMEG_0918, lacks the copper-binding motif. We generated MSMEG_0918 knockdown strains of
M. smegmatis
using CRISPRi. We confirmed the strains had varying levels of
MSMEG_0918
expression using RT-PCR. We demonstrated that MSMEG_0918 under-expression did not alter the growth of
M. smegmatis
in standard aerobic culture as compared to the wild-type. Our knockdown strains (CHROME1 and CHROME2) could be further used towards understanding the role of MSMEG_0918 in
M. smegmatis
.