TY - JOUR
T1 - Identification and functional characterization of the siRNA pathway in Taenia crassiceps by silencing Enolase A
AU - Guerrero-Hernández, Julio
AU - Bobes, Raúl J.
AU - García-Varela, Martín
AU - Castellanos-Gonzalez, Alejandro
AU - Laclette, Juan P.
N1 - Funding Information:
This report was supported in part by grants A1-5-11306 (CONACYT) and [IN 205820] PAPIIT-UNAM. JGH is a PhD student in the Doctorado en Ciencias Biológicas-UNAM and was supported by a CONACYT scholarship (No. 440780). AC and RJB were supported by CONTEX 2020-54B (UT system-Conacyt).
Publisher Copyright:
© 2021
PY - 2022/1
Y1 - 2022/1
N2 - A gene silencing procedure on cysticerci of the taeniid cestode Taenia crassiceps is described. This is the first time this technique is reported in this species that is widely used as an animal model for human cysticercosis. Genome database searches were performed in order to find out if relevant genes involved in gene silencing and non-coding RNA processing, Argonaute and Dicer (AGO and Dcr) are present in T. crassiceps. We found three AGO and two Dcr orthologues that were designed TcAGO1, Tc2 and Tc3, as well as TcDcr1 and TcDcr2. In order to elucidate the evolutionary relationships of T. crassiceps TcAGO and TcDcr genes, separate phylogenetic analyses were carried out for each, including AGO and Dcr orthologues of other 20 platyhelminthes. Our findings showed a close phylogenetic relationship of TcAGO and TcDcr with those previously described for Echinococcus spp. Our RT-PCR studies demonstrated expression of all TcAGO and TcDcr orthologues. Our results show that the gene silencing machinery in T. crassiceps is functionally active by inducing silencing of TcEnoA (∼90%). These results clearly show that gene silencing using siRNAs can be used as a molecular methodology to study gene function in taeniid cestodes.
AB - A gene silencing procedure on cysticerci of the taeniid cestode Taenia crassiceps is described. This is the first time this technique is reported in this species that is widely used as an animal model for human cysticercosis. Genome database searches were performed in order to find out if relevant genes involved in gene silencing and non-coding RNA processing, Argonaute and Dicer (AGO and Dcr) are present in T. crassiceps. We found three AGO and two Dcr orthologues that were designed TcAGO1, Tc2 and Tc3, as well as TcDcr1 and TcDcr2. In order to elucidate the evolutionary relationships of T. crassiceps TcAGO and TcDcr genes, separate phylogenetic analyses were carried out for each, including AGO and Dcr orthologues of other 20 platyhelminthes. Our findings showed a close phylogenetic relationship of TcAGO and TcDcr with those previously described for Echinococcus spp. Our RT-PCR studies demonstrated expression of all TcAGO and TcDcr orthologues. Our results show that the gene silencing machinery in T. crassiceps is functionally active by inducing silencing of TcEnoA (∼90%). These results clearly show that gene silencing using siRNAs can be used as a molecular methodology to study gene function in taeniid cestodes.
KW - Dicer and Argonaute
KW - Enolase
KW - Gene silencing
KW - Glycolysis and Krebs cycle
KW - Taenia crassiceps
KW - siRNA
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U2 - 10.1016/j.actatropica.2021.106197
DO - 10.1016/j.actatropica.2021.106197
M3 - Article
C2 - 34688628
AN - SCOPUS:85117755567
SN - 0001-706X
VL - 225
JO - Acta Tropica
JF - Acta Tropica
M1 - 106197
ER -