## 1 Introduction  
The appearance of bacterial infection is a crucial problem for burn wounds. Although there may be no infection 48 h after injury, Gram (+) bacteria, such as _Staphylococcus aureus_ and _Streptococcus_ species, may later be found in burn wounds. One week post burn Gram (-) bacteria appear to dominate. There is also an emerging antibiotic-resistant bacteria-like methicillin-resistant _Staphylococcus aureus_ (MRSA) that has been discovered in burn wounds. The increasing prevalence of MRSA poses an emerging global health threat and is associated with high rates of morbidity and mortality [1; 2; 3]. MRSA bacteremia may be treated with common topical antibiotics, such as silver sulfadiazine, neomycin, lucitacin, and muiprocin [4; 5]. Muiprocin is produced by the bacterium _Pseudomonas fluorescens_. It is highly effective against Gram-positive bacteria, such as 5. _aureus_, including MRSA. The mechanism of action is protein synthesis inhibition by binding to isoleucyl-tRNA synthetase. Muiprocin formulations are commercially available drugs used to treat skin wound infections in operative wounds, burn wounds, and diabetic wounds. Muiprocin is available as a cream and an ointment. However, the cream and ointment-basedmedications have disadvantages, such as greasiness and viscosity, which are undesirable for the drug user [6]. Moreover, 2% muiprocin ointment with polyethylene glycol requires application three times daily, leading to poor patient compliance. Furthermore, muiprocin is slowly metabolized by the skin to an inactive form. Therefore, controlled release of topical delivery of muiprocin will improve its efficacy and decrease the frequency of application to once daily [7,8]. In previous reports on muiprocin retardation of wound healing, liposomes in hydrogel [9] and nano-liposomes [10] were developed to improve therapeutics [11] and reduce toxicity compared with muiprocin alone. Nevertheless, there are limitations to the approaches, such as the sensitivity of liposomes to high temperatures during sterilization. When the temperature rises above 25 \({}^{\circ}\)C, the liposomes release the drug and other constituents [11].  
In our previous study, muiprocin nanoparticle-loaded hydrogels (MLHs) were successfully developed which were expected to enhance or improve the antibacterial activity of muiprocin [12]. Previous work on the antibacterial activity of muiprocin has primarily focused on in vitro conditions, such as minimum inhibitory concentration (MIC) and the minimum antimicrobial concentration needed to eradicate infectious organisms, or minimum biocidal concentration (MBC). These studies confirmed the antibacterial activity of muiprocin and its mechanism of action, together with gene expression analysis. To date, systematic studies of gene expression after treatment with muiprocin formulated in hydrogels have not been reported. This is probably because the bacterial gene expression purification of quantities sufficient to obtain bacterial RNA is technically difficult. RNA sequencing (RNA-seq) can offer data analysis of the entire transcriptome of a sample and can be used to investigate levels of gene expression. Gene expression analysis can be used to monitor response genes to compare the treatment of pure muiprocin with MLH. The study of gene expression can also interlace the profiles of several gene expressions to explain the similarities or differences in MRSA genes after muiprocin or MLH treatment. This technique provides a strong alternative to employing microarrays in examinations of the gene expression of MRSA. Therefore, this study investigated the inhibition of isolecyl-tRNA synthetase and global gene expression in MRSA by MLH compared with muiprocin. MRSA was stained with fluorescein diacetate (FDA) dye to monitor cell viability, and gene expression was studied via high-throughput RNA-seq. The evidence for MLH therapeutics is considered and discussed.