Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120

0537734 - MBÚ 2021 RIV US eng J - Journal Article
Srivastava, Amit - Ballal, A. - Forchhammer, K. - Tripathi, A. K.
Construction of Antisense RNA-mediated Gene Knock-down Strains in the Cyanobacterium Anabaena sp. PCC 7120.
Bio-protocol. Roč. 10, č. 4 (2020), č. článku e3528. ISSN 2331-8325
R&D Projects: GA MŠMT(CZ) LO1416
Institutional support: RVO:61388971
Keywords : AntisenseRNA (asRNA) * Knock-down * Anabaena PCC 7120 * Cyanobacteria * Green fluorescent protein * psbA1 promoter
OECD category: Microbiology
Method of publishing: Open access
https://bio-protocol.org/e3528?sid=f85bba85-5c8b-4cd0-96d2-e2c9090f9d97&sousouid=20210118012731766&ptype=0&px=1&intwzid=3528

Anabaena sp. PCC 7120 (hereafter Anabaena) is a model cyanobacterium to study nitrogen fixation, cellular differentiation and several other key biological functions that are analogous in plants. As with any other organism, many genes in Anabaena encode an essential life function and hence cannot be deleted, causing a bottleneck in the elucidation of its genomic function. Antisense RNA (asRNA) mediated approach renders the study of essential genes possible by suppressing (but not completely eliminating) expression of the target gene, thus allowing them to function to some extent. Recently, we have successfully implemented this approach using the strong endogenous promoter of the psbAl gene (D1 subunit of Photosystem II) introduced into a highcopy replicative plasmid (pAM1956) to suppress the transcript level of the target gene alr0277 (encoding a sigma factor, SigJ/Alr0277) in Anabaena. This protocol represents an efficient and easy procedure to further explore the functional genomics, expanding the scope of basic and applied research in these ecologically important cyanobacteria.
Permanent Link: http://hdl.handle.net/11104/0316052