An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum
Name
s41598-020-77644-4.pdf
Description
Published version
Size
2.93 MB
Format
Unknown
Checksum (MD5)
1d001c5ef7c67fe8ae175f320edffa6a
Author(s)
Nasamu, Armiyaw S
Falla, Alejandra
Pasaje, Charisse Flerida A
Wall, Bridget A
Wagner, Jeffrey C
Ganesan, Suresh M
Goldfless, Stephen J
Niles, Jacquin C
Date Issued
January 2021
Journal
Scientific Reports
Publisher
Springer Science and Business Media LLC
Version
Final published version
Abstract
© 2021, The Author(s). Establishing robust genome engineering methods in the malarial parasite, Plasmodium falciparum, has the potential to substantially improve the efficiency with which we gain understanding of this pathogen’s biology to propel treatment and elimination efforts. Methods for manipulating gene expression and engineering the P. falciparum genome have been validated. However, a significant barrier to fully leveraging these advances is the difficulty associated with assembling the extremely high AT content DNA constructs required for modifying the P. falciparum genome. These are frequently unstable in commonly-used circular plasmids. We address this bottleneck by devising a DNA assembly framework leveraging the improved reliability with which large AT-rich regions can be efficiently manipulated in linear plasmids. This framework integrates several key functional genetics outcomes via CRISPR/Cas9 and other methods from a common, validated framework. Overall, this molecular toolkit enables P. falciparum genetics broadly and facilitates deeper interrogation of parasite genes involved in diverse biological processes.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/S41598-020-77644-4
