Attempt of thyX gene silencing and construction of a thyX deleted clone in a Mycobacterium bovis BCG

Mycobacterium tuberculosis, the causative agent of tuberculosis, possess flavin-dependent thymidylate synthase, ThyX. Since thyX is absent in humans and was shown to be essential for M. tuberculosis normal growth, ThyX is thought to be an attractive novel TB drug target. This study assessed thyX essentiality in Mycobacterium bovis BCG strains using CRISPR interference based gene silencing and found that thyX is not essential in an M. bovis BCG Tokyo derivative strain. A thyX deletion mutant strain was successfully constructed from that strain, which reinforces the non-essentiality of thyX under a certain genetic background.

This is the peer reviewed version of the following article: [Arimura, Y, Minato, Y, Wada, T, Nakayama, M, Ryumon, A, Hirata, N, et al. Attempt of thyX gene silencing and construction of a thyX deleted clone in a Mycobacterium bovis BCG. Microbiol Immunol. 2021; 1– 5. https://doi.org/10.1111/1348-0421.12944], which has been published in final form at [https://doi.org/10.1111/1348-0421.12944]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages there of by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.