Forward Genetics in African Trypanosomes.

Methods Mol Biol

Trypanosome Molecular Biology Group, Institut Pasteur, Paris, France.

Published: February 2021

Forward genetic screens in Trypanosoma brucei have enabled researchers to move from a candidate-gene based approach to one where we are able to studying all genes required for a single process simultaneously. In this protocol, we describe how to generate RNAi library strains in bloodstream form trypanosomes, run a screen by selecting for drug resistance or using a reporter gene and process the high-throughput sequencing data for a genome scale RNAi library screen.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-0294-2_21DOI Listing

Publication Analysis

Top Keywords

rnai library
8
forward genetics
4
genetics african
4
african trypanosomes
4
trypanosomes forward
4
forward genetic
4
genetic screens
4
screens trypanosoma
4
trypanosoma brucei
4
brucei enabled
4

Similar Publications

Targeting NUCKS1 with a fragment of tRNA of Chinese yew for the treatment of colorectal cancer.

Noncoding RNA Res

April 2025

State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Avenida Wai Long, Taipa, Macau SAR, China.

Despite the discovery of numerous oncogenes in colorectal cancer (CRC), the development of associated drugs is limited, posing a significant challenge for CRC treatment. Identification of novel druggable targets is therefore crucial for the therapeutic development of CRC. Here, we report the first investigation on therapeutics targeting the potent oncogene NUCKS1 to suppress cancer progression.

View Article and Find Full Text PDF

RBM19 promotes the progression of prostate cancer under docetaxel treatment via SNHG21/PIM1 axis.

Cell Biol Toxicol

December 2024

Department of Urology, Jinjiang Municipal Hospital, Luoshan Section, No. 16 Jinguang Road, Luoshan Street, Jinjiang City, Quanzhou, Fujian, China.

RBM family proteins plays the critical role in the progression of numerous tumors. However, whether RBM family proteins involved in prostate cancer (PCa) progression is remain elucidated. In our study, an RNAi screen containing shRNA library targeting 54 members of the RBM family was applied to identify the critical RBM proteins involved in prostate cancer progression under docetaxel treatment, and RBM19 was selected.

View Article and Find Full Text PDF

RNAi library screening reveals Gβ1, Casein Kinase 2 and ICAP-1 as novel regulators of LFA-1-mediated T cell polarity and migration.

Immunol Cell Biol

January 2025

Trinity Translational Medicine Institute & Department of Clinical Medicine, Trinity College Dublin, St James's Hospital, Dublin, Republic of Ireland.

The αβ integrin LFA-1 plays a key role in T-cell adhesion to the endothelial vasculature and migration into both secondary lymphoid organs and peripheral tissues via interactions with its target protein ICAM-1, but the pathways that regulate LFA-1-mediated T-cell polarity and migration are not fully understood. In this study we screened two RNAi libraries targeting G protein-coupled receptors (GPCR)/GPCR-associated proteins and kinases in a HuT 78 T cell line model of LFA-1-stimulated T-cell migration. Based on staining of the actin cytoskeleton, multiple parameters to measure cell morphology were used to assess the contribution of 1109 genes to LFA-1-mediated T-cell polarity and migration.

View Article and Find Full Text PDF

RNAi Screen Identifies AXL Inhibition Combined with Cannabinoid WIN55212-2 as a Potential Strategy for Cancer Treatment.

Pharmaceuticals (Basel)

November 2024

State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.

: Cannabinoids are commonly used as adjuvant cancer drugs to overcome numerous adverse side effects for patients. The aim of this study was to identify the target genes that show a synergistic anti-tumor role in combination with the cannabinoid WIN55212-2 in vitro and in vivo. : A human kinome RNAi library was used to screen the targeted gene that silencing plus WIN55212-2 treatment synergistically inhibited cancer cell growth in an INCELL Analyzer 2000.

View Article and Find Full Text PDF

RNAi therapeutics possess the potential to cure many uncurable human diseases. For instance, RNAi therapeutics using liposomes showed remarkable survival benefits in patients with liver diseases. However, the extension of liposomes to deliver RNA to cure other ailments has largely been unsuccessful.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!