In this section
@article{Budzak:2023:10.1016/j.molbiopara.2023.111598,
author = {Budzak, J and Goodwin, I and Tiengwe, C and Rudenko, G},
doi = {10.1016/j.molbiopara.2023.111598},
journal = {Molecular and Biochemical Parasitology},
title = {Imaging of genomic loci in Trypanosoma brucei using an optimised LacO-LacI system},
url = {http://dx.doi.org/10.1016/j.molbiopara.2023.111598},
volume = {256},
year = {2023}
}
TY - JOUR
AB - Visualisation of genomic loci by microscopy is essential for understanding nuclear organisation, particularly at the single cell level. One powerful technique for studying the positioning of genomic loci is through the Lac Operator-Lac Repressor (LacO-LacI) system, in which LacO repeats introduced into a specific genomic locus can be visualised through expression of a LacI-protein fused to a fluorescent tag. First utilised in Trypanosoma brucei over 20 years ago, we have now optimised this system with short, stabilised LacO repeats of less than 2 kb paired with a constitutively expressed mNeongreen::LacI fusion protein to facilitate visualisation of genomic loci. We demonstrate the compatibility of this system with super-resolution microscopy and propose its suitability for multiplexing with inducible RNAi or protein over expression which will allow analysis of nuclear organisation after perturbation of gene expression.
AU - Budzak,J
AU - Goodwin,I
AU - Tiengwe,C
AU - Rudenko,G
DO - 10.1016/j.molbiopara.2023.111598
PY - 2023///
SN - 0166-6851
TI - Imaging of genomic loci in Trypanosoma brucei using an optimised LacO-LacI system
T2 - Molecular and Biochemical Parasitology
UR - http://dx.doi.org/10.1016/j.molbiopara.2023.111598
UR - https://www.ncbi.nlm.nih.gov/pubmed/37923299
UR - https://www.sciencedirect.com/science/article/pii/S0166685123000567
UR - http://hdl.handle.net/10044/1/108307
VL - 256
ER -
Dr. Calvin Tiengwe
Dept. of Life Sciences,
503 Sir Ernst Chain Building,
Imperial College London,
SW7 2AZ, UK
