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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

A suitable RNA preparation methodology for whole transcriptome shotgun sequencing harvested from Plasmodium vivax-infected patients

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Autor(es):
Bourgard, Catarina [1] ; Lopes, Stefanie C. P. [2, 3] ; Lacerda, Marcus V. G. [2, 3] ; Albrecht, Letusa [1, 4] ; Costa, Fabio T. M. [1]
Número total de Autores: 5
Afiliação do(s) autor(es):
[1] Univ Campinas UNICAMP, Inst Biol, Dept Genet Evolut Microbiol & Immunol, Lab Trop Dis, Campinas, SP - Brazil
[2] Fundacao Oswaldo Cruz FIOCRUZ, Inst Leonidas & Maria Deane, Manaus, AM - Brazil
[3] Fundacao Med Trop Dr Heitor Vieira Dourado FMT HV, Gerencia Malaria, Manaus, AM - Brazil
[4] Fundacao Oswaldo Cruz Fiocruz, Inst Carlos, Curitiba, PR - Brazil
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: SCIENTIFIC REPORTS; v. 11, n. 1 MAR 3 2021.
Citações Web of Science: 1
Resumo

Plasmodium vivax is a world-threatening human malaria parasite, whose biology remains elusive. The unavailability of in vitro culture, and the difficulties in getting a high number of pure parasites makes RNA isolation in quantity and quality a challenge. Here, a methodological outline for RNA-seq from P. vivax isolates with low parasitemia is presented, combining parasite maturation and enrichment with efficient RNA extraction, yielding similar to 100 pg.mu L-1 of RNA, suitable for SMART-Seq Ultra-Low Input RNA library and Illumina sequencing. Unbiased coding transcriptome of similar to 4 M reads was achieved for four patient isolates with similar to 51% of transcripts mapped to the P. vivax P01 reference genome, presenting heterogeneous profiles of expression among individual isolates. Amongst the most transcribed genes in all isolates, a parasite-staged mixed repertoire of conserved parasite metabolic, membrane and exported proteins was observed. Still, a quarter of transcribed genes remain functionally uncharacterized. In parallel, a P. falciparum Brazilian isolate was also analyzed and 57% of its transcripts mapped against IT genome. Comparison of transcriptomes of the two species revealed a common trophozoite-staged expression profile, with several homologous genes being expressed. Collectively, these results will positively impact vivax research improving knowledge of P. vivax biology. (AU)

Processo FAPESP: 12/16525-2 - Plasmodium vivax: patogênese e infectividade
Beneficiário:Fabio Trindade Maranhão Costa
Linha de fomento: Auxílio à Pesquisa - Temático
Processo FAPESP: 13/20509-5 - Análise dos mecanismos imunopatológicos e moleculares envolvidos no processo de citoaderência de Plasmodium vivax
Beneficiário:Catarina Baeta da Luz Bourgard
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 17/18611-7 - Desenvolvimento de novas ferramentas para busca e validação de alvos moleculares para terapia contra Plasmodium vivax
Beneficiário:Fabio Trindade Maranhão Costa
Linha de fomento: Auxílio à Pesquisa - Temático