Abstract
Leishmania is the causative agent of leishmaniasis, a neglected tropical disease that affects more than 12 million people around the world. Current treatments are toxic and poorly effective due to the acquisition of resistance within Leishmania populations. Thus, the pursuit for new antileishmanial drugs is a priority. The available methods for drug screening based on colorimetric assays using vital dyes are time-consuming. Currently, the use of fluorescent reporter proteins is replacing the use of viability indicator dyes. We have constructed two plasmids expressing the red fluorescent protein mCherry with multiple cloning sites (MCS), adequate for N- and C-terminal fusion protein constructs. Our results also show that the improved pXG-mCherry plasmid can be employed for drug screening in vitro. The use of the red fluorescent protein, mCherry, is an easier tool for numerous assays, not only to test pharmacological compounds, but also to determine the subcellular localization of proteins.
Highlights
Leishmaniasis is a neglected tropical disease caused by protozoans of the genus Leishmania
According to the World Health Organization (WHO), 350 million people are at risk of infection as no commercial vaccines are available for its prevention [1]
A DNA region containing the coding sequence for mCherry with different multiple cloning sites (MCS) was inserted into the Leishmania expression vector pXG, generating two pXG-mCherry vectors
Summary
Leishmaniasis is a neglected tropical disease caused by protozoans of the genus Leishmania. Since the recognition of Leishmania spp. as the causative agent of leishmaniasis, generic pentavalent antimonials have been the essential chemotherapy agents against this disease. Pentostam and Glucantime are the branded alternatives to generic antimonials treatment. Other treatments such as amphotericin B and miltefosine offer a higher efficacy but a costlier option than pentavalent antimonials. Antipathogens like paromomycin and pentamidine show some benefit in the treatment of leishmaniasis, especially when used in conjunction with other drugs. All these pharmacological compounds raise severe side effects without the confidence of complete healing and currently, and various Leishmania strains have developed resistance against these drugs [2]
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