Patent application EP19382445A (EP), filed 2019-05-31
Publication number: EP3744849A1
Title: METHOD FOR MODULATING THE GENE SILENCING DEGREE INDUCED BY TRANS-ACTING SMALL INTERFERING RNAS
Abstract: The invention provides a method for fine-tuning the gene silencing degree induced by a synthetic trans-acting small interfering RNA (syn-tasiRNA) which comprises varying the insertion position of the sequence of the syn-tasiRNA in its TAS1c precursor, regarding the miR173 cleavage target site, in order to obtain a higher or lower amount of the syn-tasiRNA expressed and consequently a higher or lower silencing activity induced by the same.
IPC codes: C12N  15/    82            A I
Claims: number: 1
text: A method for modulating the gene silencing degree induced by a synthetic trans-acting small interfering RNA over at least one target gene in a subject, wherein said method comprises the following steps:
number: 2
text: The method according to claim 1 for modulating the gene silencing degree of two different target genes by two different synthetic trans-acting small interfering RNAs, wherein said method comprises, in step (b), inserting two different sequences of the synthetic trans-acting small interfering RNAs at different positions of the transcript of step (a) depending on the gene silencing degree desired for each target gene.
number: 3
text: The method according to any of claims 1 or 2, wherein the subject is a plant.
number: 4
text: The method according to claim 3, wherein the plant is selected from the list consisting of cotton, potato, corn, wheat, rice, sugar cane, oilseed rape, Arabidopsis thaliana, Nicotiana benthamiana, sugarbeet, tobacco and soybean.
number: 5
text: The method according to claim 4, wherein the plant is Arabidopsis thaliana or Nicotiana benthamiana.
number: 6
text: The method according to any of claims 1 to 5, wherein the synthetic trans-acting small interfering RNA is a synthetic trans-acting small interfering RNA against a target gene selected from the list consisting of: FT, CH42 or TSWV.
number: 7
text: The method according to any of claims 1 to 6, wherein the subject is an Arabidopsis thaliana plant and the target gene is FT or CH42.
number: 8
text: The method according to any of claims 1 to 6, wherein the subject is a Nicotiana benthamiana plant and the target is TSWV.
number: 9
text: The method according to any of claims 1 to 8, wherein the position near the miR173 cleavage target site in the transcript is D2.
number: 10
text: The method according to any of claims 1 to 9, wherein the position far away from the miR173 cleavage target site in the transcript is D4 or D5.
number: 11
text: The method according to claim 10, wherein the position far away from the miR173 cleavage target site in the transcript is D5.
Office action: search_report, dated 2020-12-02
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ref_id: - ALBERTO CARBONELL, "Secondary Small Interfering RNA-Based Silencing Tools in Plants: An Update", FRONTIERS IN PLANT SCIENCE, (20190528), vol. 10, doi:10.3389/fpls.2019.00687, XP055639161 [I] 1-11 * abstract; pages 1-4; Figure 1A *
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ref_id: - A. CARBONELL ET AL, "New Generation of Artificial MicroRNA and Synthetic Trans-Acting Small Interfering RNA Vectors for Efficient Gene Silencing in Arabidopsis", PLANT PHYSIOLOGY, (20140319), vol. 165, no. 1, doi:10.1104/pp.113.234989, ISSN 0032-0889, pages 15 - 29, XP055123679 [I] 1-11 * abstract; pages 22, 25 and 26; Figures 6 and 7 *
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ref_id: - TAIOWA A MONTGOMERY ET AL, "AGO1-miR173 complex initiates phased siRNA formation in plants", vol. 105, no. 51, doi:10.1073/PNAS.0810241105, ISSN 0027-8424, (20081201), pages 20055 - 20062, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES (PNAS), NATIONAL ACADEMY OF SCIENCES, US, URL: http://www.pnas.org/content/105/51/20055, (20191106), XP002712275 [I] 1-11 * abstract; pages 20055, 20056, 20061, 20062; Figures 1, 2 and 4 *
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severity: background
ref_id: - Alberto Carbonell, "Design and High-Throughput Generation of Artificial Small RNA Constructs for Plants : Methods and Protocols", Methods in Molecular Biology, US, Humana Press, New York, NY, (20190101), vol. 1932, pages 247 - 260, doi:10.1007/978-1-4939-9042-9_19, ISSN 1064-3745, XP055638897 [I] 1-11 * Sections 1 (Introduction), 3.2 and 4 (in particular items 18, 20 and 21) *
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Final disposition: pending (2020-12-02)