This study shows the successfully targeted manipulation of the L. In addition, the parasite count was significantly different in the case and control of infected macrophages (P < 0.05). major creates a new stop codon and disrupts the frame sheet of the gene by creating a new insertion (thymine), which prevents its expression. Our data validate that the use of CRISPR/Cas9 in L. major was investigated by counting phagocytic parasites into macrophages and DNA sequence analysis. The effect of CRISPR/Cas9 constructs on GP63 mutation, viability, and status of L. major parasites were transfected with CRISPR/Cas9 vectors constructed by electroporation and then added to macrophage cells on RPMI. major pathogenicity by manipulating the GP63 gene. Therefore, our main goal was to evaluate the CRISPR PX-LmGP63 vector effect on pathogenicity of Leishmania major in vitroto challenge for using CRISPR/Cas9 as a therapeutic CL through the reduction of L. CRISPR/Cas9 as a powerful genome-editing tool provides the opportunity to create precise genetic manipulation to investigate the molecular basis of different leishmaniasis cases. It is, therefore, essential to find new medicines to treat leishmaniasis. The current treatment approaches are limited by the complications such as loss of fertility and drug resistance. Despite the high incidence and prevalence of Leishmania cases in many countries, it has been a neglected tropical disease. These findings provide further incentive to recapitulate the legitimacy of current control measures and resolve dynamics of resistance in human head louse populations.Ĭutaneous leishmaniasis (CL) is caused by intracellular obligate parasites (Leishmania spp.) carried by the blood-sucking of female sandflies and transmitted between mammalian hosts. The detection of six amino acid substitutions from 30% of examined head lice among infested schoolgirls reveal that mutants are minutely developing. Other exon or intron regions remained non-mutated from the remaining seven counties. Single (L840F), double (I836L, E837K), and triple novel point mutations (V875L, Q876P, S879V) the last involving two concomitant allelic substitutions were discovered in the second and third exon regions of head louse DNA on chromosome II from three (30%) counties. Sequence analysis showed that the sodium channel genes in the pooled ectoparasites had two intron and three exon regions. Samples were investigated by polymerase chain reactions (PCR), and the amplicon was subsequently sequenced. Following species identification with valid entomological keys, 350 (68%) out of 514 randomly collected adult head lice were analyzed after their somatic genomic DNA extraction using Sinaclon kit.
The specimens were screened in 10 pools (each pool per county containing 35 specimens), with three pools (30%) being positive. Head lice were collected using plastic detection combs on girls enrolled in public schools from 10 counties in Fars province. The aim of this study was to investigate kdr gene mutations on voltage-sensitive sodium channel (VSSC) among wild head lice samples from Fars province, southern Iran. Molecular characterization of this resistance has a crucial impact on selecting appropriate treatment protocol. Reduced sensitivity to treatment due to genetic mutations, in particular knockdown resistance (kdr) (or target site insensitivity) allele, has led to this infestation prevalence. The PiggyBac transposase coding sequence has been optimized for high expression, stability and activity in mammalian cells.Human head louse, Pediculus humanus capitis De Geer, 1767 (Anoplura: Pediculidae), is one of the most frequent ectoparasites infesting Homo sapiens worldwide. The powerful activity of the piggyBac transposon system enables genes of interest between the two ITRs in the PB vector to be easily mobilized into target genomes.The Super PiggyBac transposase expression vector features a 5’ HS4 insulator to support robust transcription from the rPolr2A promoter. During transposition, the PB transposase recognizes transposonspecific inverted terminal repeat sequences (ITRs) located on both ends of the transposon vector and efficiently moves the contents from the original sites and efficiently integrates them into TTAA chromosomal sites. Super PiggyBac Transposase (PB200PA-1) is a Mammalian cell carrier.The PiggyBac (PB) transposon is a mobile genetic element that efficiently transposes between vectors andchromosomes via a "cut and paste" mechanism. Super PiggyBac Transposase (PB200PA-1) Information