X-Meeting presentations

ASSEMBLY, ANNOTATION AND GENE EDITING OF THE GENOME OF THE PH8 STRAIN OF LEISHMANIA AMAZONENSIS WITH FOCUS ON MULTIGENE FAMILIES ENCODING VIRULENCE FACTORS

Published in 26/04/2022 - ISBN: 978-65-5941-645-5

Link All Papers
LinkSubmission
Paper Title
ASSEMBLY, ANNOTATION AND GENE EDITING OF THE GENOME OF THE PH8 STRAIN OF LEISHMANIA AMAZONENSIS WITH FOCUS ON MULTIGENE FAMILIES ENCODING VIRULENCE FACTORS
Authors
  • Wanessa Moreira Goes
  • Viviane Grazielle da Silva
  • Carlos Rodolpho Ferreira Brasil
  • Bárbara Emmanuelle Lacerda de Moura
  • João Luís Reis Cunha
  • Anderson Coqueiro-dos-Santos
  • Daniella Bartholomeu
  • Ana Paula Salles Moura Fernandes
  • Santuza Maria Ribeiro Teixeira
Modality
Xpress presentation
Subject area
Omics
Publishing Date
26/04/2022
Country of Publishing
Brasil
Language of Publishing
Inglês
Paper Page
https://www.even3.com.br/anais/xmeetingxp2021/423243-assembly-annotation-and-gene-editing-of-the-genome-of-the-ph8-strain-of-leishmania-amazonensis-with-focus-on-mul
ISBN
978-65-5941-645-5
Keywords
Leishmania amazonensis, annotation, multigene families
Summary
Leishmania amazonensis is one of the etiological agents of cutaneous leishmaniasis. Parasite virulence factors have already been studied, as they play a crucial role in the establishment of infection and disease development in the mammalian host. We report the sequencing and assembly of the PH8 strain of L. amazonenis combining long PacBio reads, short Illumina reads and synteny data with the Leishmania mexicana genome. The final assembly, composed of 34 chromosomes and 44 scaffolds not incorporated, represents a genome of ~ 32 Mb. The annotation of the L. amazonensis genome was transferred from the annotation of 8225 genes present in the genome of L. mexicana. Multigene families, such as amastins, metalloproteins GP63, A2 protein, cysteine proteases, fatty acid synthases, phosphatase and kinases, were identified using an automated pipeline, in which proteins from others Leishmania species and trypanosomatids interrogated. In addition, amastins and GP63 were characterized. In total, 25 genes encoding amastins were predicted in L. amazonensis. The alignment of this genes, using M-Coffee and phylogeny analyzes, using the maximum-likelihood estimation resulted in groupings corresponding to the four sub-classes of amastins (a, ß, ? and d). Analysis of the 11 sequences encoding GP63 showed the conservation of important domains, such as HExxH and SRYD, which are important for protein structure and binding to macrophage surface receptors, respectively. Finally, we tested a CRISPR-Cas9 protocol to generate knockout cell lines of the Miltefosine Transporter (TM) gene of L. amazonensis as a proof of concept. Expression of Streptococcus pyogenes Cas9 in promastigotes was achieved after transfection with pLDCN, an episomal vector. Alternatively, promastigotes were transfected with recombinant Staphylococcus aureus Cas9 ribonucleoprotein complex and an sgRNA. Both strategies were able to disrupt the TM gene in transfected parasites, originating knockout parasite cell lines.
Title of the Event
X-Meeting XPerience 2021
Title of the Proceedings of the event
X-Meeting presentations
Name of the Publisher
Even3
Means of Dissemination
Meio Digital

How to cite

GOES, Wanessa Moreira et al.. ASSEMBLY, ANNOTATION AND GENE EDITING OF THE GENOME OF THE PH8 STRAIN OF LEISHMANIA AMAZONENSIS WITH FOCUS ON MULTIGENE FAMILIES ENCODING VIRULENCE FACTORS.. In: X-Meeting presentations. Anais...São Paulo(SP) AB3C, 2021. Available in: https//www.even3.com.br/anais/xmeetingxp2021/423243-ASSEMBLY-ANNOTATION-AND-GENE-EDITING-OF-THE-GENOME-OF-THE-PH8-STRAIN-OF-LEISHMANIA-AMAZONENSIS-WITH-FOCUS-ON-MUL. Access in: 04/08/2025

Paper

Even3 Publicacoes