Reprogramming CRISPR-Mediated RNA Interference for Silencing of Essential Genes in Sulfolobales

Autor(en)
Erika Wimmer, Isabelle Anna Zink, Christa Schleper
Abstrakt

The manipulation of gene expression levels in vivo is often key to elucidating gene function and regulatory network interactions, especially when it comes to the investigation of essential genes that cannot be deleted from the model organism’s genome. Several techniques have been developed for prokaryotes that allow to interfere with transcription initiation of specific genes by blocking or modifying promoter regions. However, a tool functionally similar to RNAi used in eukaryotes to efficiently degrade mRNA posttranscriptionally did not exist until recently. Type III CRISPR-Cas systems use small RNAs (crRNAs) that guide effector complexes (encoded by cas genes) which act as site-specific RNA endonuclease and can thus be harnessed for targeted posttranscriptional gene silencing. Guide RNAs complementary to the desired target mRNA that, in addition, exhibit complementarity to repeat sequences found in the CRISPR arrays, effectively suppress unspecific DNA and RNA activities of the CRISPR-Cas complexes. Here we describe the use of endogenous type III CRISPR-Cas systems in two model organisms of Crenarchaeota, Saccharolobus solfataricus and Sulfolobus acidocaldarius.

Organisation(en)
Department für Funktionelle und Evolutionäre Ökologie
Externe Organisation(en)
Wageningen University and Research Centre
Band
2522
Seiten
177-201
Anzahl der Seiten
25
DOI
https://doi.org/10.1007/978-1-0716-2445-6_11
Publikationsdatum
09-2022
Peer-reviewed
Ja
ÖFOS 2012
106022 Mikrobiologie
Schlagwörter
ASJC Scopus Sachgebiete
Genetics, Molecular Biology
Link zum Portal
https://ucris.univie.ac.at/portal/de/publications/reprogramming-crisprmediated-rna-interference-for-silencing-of-essential-genes-in-sulfolobales(50066b07-b5c5-4019-80f6-0691c9d49cbf).html