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Trypanosoma brucei brucei Doxycycline induced DRBD18 knockdown whole cell RNA and cytoplasmic RNA

ABSTRACT: Trypanosoma brucei brucei Doxycycline induced DRBD18 knockdown whole cell RNA and cytoplasmic RNA

PROVIDER: PRJNA665716 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12712967_1.fastq.gz	Fastqsanger.gz
	SRR12712967_2.fastq.gz	Fastqsanger.gz
	SRR12712968_1.fastq.gz	Fastqsanger.gz
	SRR12712968_2.fastq.gz	Fastqsanger.gz
	SRR12712969_1.fastq.gz	Fastqsanger.gz

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Trypanosoma brucei brucei Doxycycline induced DRBD18 knockdown whole cell RNA and cytoplasmic RNA

Project description:DRBD18 (Tb927.11.14090) was knocked down by 19 hour doxycycline treatment. Whole cell RNA and cytoplasmic RNA was isolated and sequenced.

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Project description:In this work, we examine the function of DRBD18 in T. brucei, here using single cell RNA sequencing (scRNAseq) of uninduced and induced DRBD18 knockdowns. In insect procyclic form, we identified one major and one minor population of cells that dramatically increased upon DRBD18 knockdown. Both of these cell populations were significantly enriched for transcripts that are normally upregulated in the quiescent stumpy bloodstream form and metacyclic (MF) life cycle stages, consistent with a role for DRBD18 in life cycle progression. Furthermore, the 3’ end bias of our sequencing technique allowed us to investigate the effect of DRBD18 on poly(A) site selection, and extension of gene annotations permitted a comprehensive analysis of these changes. We found that 3’UTR shortening upon DRBD18 depletion was prevalent, with over 1500 transcripts exhibiting this this phenotype, and RIPseq analyses showed significant overlap between bound transcripts and those with 3’UTR shortening

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RNAi against DRBD18 in Trypanosoma brucei bloodstream forms

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In vitro generation of human high-density-lipoprotein-resistant Trypanosoma brucei brucei

Project description:The host range of African trypanosomes is influenced by innate protective molecules in the blood of primates. A subfraction of human high-density lipoprotein (HDL) containing apolipoprotein A-I, apolipoprotein L-I, and haptoglobin-related protein is toxic to Trypanosoma brucei brucei but not the human sleeping sickness parasite Trypanosoma brucei rhodesiense. It is thought that T. b. rhodesiense evolved from a T. b. brucei-like ancestor and expresses a defense protein that ablates the antitrypanosomal activity of human HDL. To directly investigate this possibility, we developed an in vitro selection to generate human HDL-resistant T. b. brucei. Here we show that conversion of T. b. brucei from human HDL sensitive to resistant correlates with changes in the expression of the variant surface glycoprotein (VSG) and abolished uptake of the cytotoxic human HDLs. Complete transcriptome analysis of the HDL-susceptible and -resistant trypanosomes confirmed that VSG switching had occurred but failed to reveal the expression of other genes specifically associated with human HDL resistance, including the serum resistance-associated gene (SRA) of T. b. rhodesiense. In addition, we found that while the original active expression site was still utilized, expression of three expression site-associated genes (ESAG) was altered in the HDL-resistant trypanosomes. These findings demonstrate that resistance to human HDLs can be acquired by T. b. brucei. Keywords: Trypanosoma, VSG, antigenic switching, HDL-resistance

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