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Method for increasing cannabis sativa yield by gene editing

A hemp, yield technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., to solve problems such as unmet demand

Pending Publication Date: 2022-05-06
BETTERSEEDS LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Consequently, there has been a long felt and unmet need to provide cannabis strains with improved yields

Method used

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  • Method for increasing cannabis sativa yield by gene editing
  • Method for increasing cannabis sativa yield by gene editing
  • Method for increasing cannabis sativa yield by gene editing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Example 1: General scheme of the method for generating genome-edited plants

[0084] now refer to Figures 1 to 3 , which discloses methods of generating genome-edited cannabis plants. Various common cannabis tissues ([A] auxiliary buds; [B] mature leaves; [C] callus; [D] cotyledons, such as figure 1 shown) for conversion. For successful transformation, use the following protocol:

[0085] 1. Design and synthesis of gRNA corresponding to the targeting sequence for editing. Editing events should be designed flanked by unique restriction site sequences to allow easier screening for successful edits.

[0086] 2. Transformation using Agrobacterium or bioballistics. For Agrobacterium and biolistics using DNA plasmids, construct a vector containing the selectable marker, Cas9 gene, and associated gRNA. For bioballistics using ribonucleoprotein (RNP) complexes, the RNP complexes are generated by mixing the Cas9 protein with the relevant gRNA.

[0087] 3. Regeneration in...

example 2

[0095] Example 2: gRNA sequences of common cannabis genes disclosed in this application

[0096] Reference is now made to the following tables which present non-binding examples of gRNA sequences for common cannabis genes disclosed in this application, along with their respective positions, strands and PAMs (protospacer adjacent motifs).

[0097] Table 1: gRNA (guide RNA) sequence and complementary PAM (protospacer adjacent motif) of CsBif1 (referred to in the sequence listing file as SEQ ID NO: 4-170).

[0098]

[0099]

[0100]

[0101]

[0102] Location chain sequence PAM 1334 1 TAGATGGTAAAAATAAACAA AGG 1352 1 AAAGGAAAGCTGTATCTAAT TGG 1353 1 AAGGAAAGCTGTATCTAATT GGG 1373 -1 TAAAAGAACTTTTGGAACTG TGG 1381 -1 TTTAATGCTAAAAGAACTTT TGG 1406 1 AGCATTAAATGAAGAAAAAT TGG 1415 1 TGAAGAAAAATTGGCAAAGA TGG 1421 1 AAAATTGGCAAAGATGGAAA AGG 1441 1 AGGTCTCAATAGTTGAAATT TGG 1510 -...

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Abstract

A method for increasing flower yield in cannabis sativa plants by a genome editing method is disclosed. More specifically, the method comprises the steps of: (a) selecting a gene involved in the flowering pathway of the Cannabis sativa species; (b) synthesizing or designing gRNA expression cassettes corresponding to targeted cleavage loci along the hemp genome; (c) transforming the cannabis plant cells to insert genetic material therein; (d) culturing the cannabis plant cells; (e) selecting the cannabis cell expressing the desired mutation at the editing target region, and (f) regenerating a plant from the transformed plant cell, plant nucleus or plant tissue.

Description

[0001] sequence listing [0002] This application contains a Sequence Listing filed electronically in ASCII txt. format, which is hereby incorporated by reference in its entirety. The ASCII copy is named Sequence Listing 2272-C-01-PCT and is 457KB in size. technical field [0003] The present invention relates generally to the field of improving plant traits. More specifically, the present invention relates to increasing flower yield in cannabis plants using the CRISPR / Cas genome editing approach. Background technique [0004] The marijuana market is enjoying an unprecedented surge in activity following the widespread legalization trend across the globe. The U.S. market alone is estimated to be worth at least $30 billion by 2025, growing at a rare rate of 30% per year. This has led to increased demand not only for cannabis products in general, but especially for products with very specific traits, whether for medicinal or recreational use. That demand is sometimes met wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/55A01H5/10A01H5/02A01H6/28
CPCA01H5/02C12N15/8213C12N15/827Y02A40/146C12N2310/20C12N9/22C12N15/11
Inventor 伊多·马葛利特塔尔·谢尔曼希拉·科里姆
Owner BETTERSEEDS LTD