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2014 | 36 | 08 |

Tytuł artykułu

Design of artificial miRNA for redundant silencing of Brassica SHP1 and SHP2: transient assay-based validation of transcript cleavage from polyploid Brassicas

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
Asynchronous pod-shatter results in significant yield losses in canola. SHATTERPROOF1 (SHP1) and SHATTERPROOF2 (SHP2) regulate pod shatter in Brassicaceae. This study describes the potential of an artificial RNA (amiRNA)-based approach for redundant silencing of SHP1/SHP2 homologs in Brassicas for introducing shatter resistance. After surveying the natural variation in Brassica SHP homologs, amiR-bnashp1 and amiR-bnashp2 were designed and processed from miR319a precursor in N. benthamiana. Analysis of thermodynamic interaction between amiRNAs and SHP1/SHP2 homologs predicted high binding energy for amiR-bnashp2 [total free energy of binding (TFEB): -23.54 kcal/mol] and not for amiR-bnashp1 (TFEB: 6.01 kcal/mol). As predicted, agroco-infiltration assay validated amiR-bnashp2-mediated cleavage of SHP1 and SHP2 homologs from B. napus, B. juncea and A. thaliana, while amiR-bnashp1 failed to direct cleavage of these homologs. In conclusion, for trait manipulation in polyploid genomes deployment of amiRNAs is suggested to silence redundant genes. Furthermore, an a priori knowledge on transcriptome potential of genome concerned is a prerequisite for predicting efficiency and specificity of target selection by amiRNA(s).

Słowa kluczowe

Wydawca

-

Rocznik

Tom

36

Numer

08

Opis fizyczny

p.2125-2135,fig.,ref.

Twórcy

autor
  • Department of Biotechnology, TERI University, 10 Institutional Area, Vasant Kunj, 110070 New Delhi, India
  • Department of Biotechnology, TERI University, 10 Institutional Area, Vasant Kunj, 110070 New Delhi, India
autor
  • Department of Biotechnology, TERI University, 10 Institutional Area, Vasant Kunj, 110070 New Delhi, India

Bibliografia

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Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

bwmeta1.element.agro-a6fb54e7-7fc3-4dc1-86da-5a37dbfa8c68
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