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Living Modified Organism
(LMO)
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Maize with altered growth characteristics
EN
B104 (NGAL2-KO)
No
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Person:Dr. Hilde Nelissen, Departement PlantensysteembiologieTechnologiepark 927Gent,
B-9052, BelgiumPhone: +32 9 2446611,Fax:Email: vib@vib.be,Website: http://www.vib.be,Related OrganizationVIB - Universiteit Gent (VIB-UGent)Academic or research instituteTechnologiepark 927Gent,
B-9052, BelgiumPhone: +32 9 2446611,Fax:Email: vib@vib.be,Website: http://www.vib.be,
The maize plants have been edited using the CRISPR/Cas9 system resulting in the mutation of the NGAL2 gene. One extra DNA base pair has been introduced into the target gene resulting in the knock-out of the gene by means of a frameshift mutation.
The NGAL2 is a transcription factor that has been shown to negatively affect the expression of the PLA1 gene. The PLA1 gene codes for a cytochrome P450 mono-oxygenase enzyme that is involved in the production of factors that control cell proliferation. Due to the removal of the transcriptional repressor, increased expression of PLA1 is expected and would alter growth characteristics, which are phenotypically shown by resulting in a significant larger leaf size and increased biomass.
EN
The NGAL2 is a transcription factor that has been shown to negatively affect the expression of the PLA1 gene. The PLA1 gene codes for a cytochrome P450 mono-oxygenase enzyme that is involved in the production of factors that control cell proliferation. Due to the removal of the transcriptional repressor, increased expression of PLA1 is expected and would alter growth characteristics, which are phenotypically shown by resulting in a significant larger leaf size and increased biomass.
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
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BCH-ORGA-SCBD-246-6 Organism Zea mays (Maize, Corn, MAIZE)Crops
Zea Mays inbred line B104
EN
pBUN411
EN
- Agrobacterium-mediated DNA transfer
- Other (CRISPR-Cas9 technique)
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-115122-1 NGAL2 transcription factor | Zea mays (Maize, Corn, MAIZE)Protein coding sequence | Changes in physiology and/or production (Yield)
Utilizing Agrobacterium tumefaciens-mediated transformation, a gene cassette containing the CRISPR-Cas9 machinery was inserted into the genome. The gene editing machinery introduced a single basepair insertion into the coding sequence of NGAL2 through a DNA repair mechanism and resulted in a frameshift mutation. Transcription is under control of the native promoter and terminator.
Transformants containing the gene cassette and the desired mutations were selected, and by means of conventional crossing with wild-type plants, the CRISPR/Cas9 gene cassette was removed by selecting T1 plants that only contained the desired mutation, but no longer contained the gene cassette (null-segregants). Therefore, the plants in the field trial are not expected to contain foreign genetic material. They should only contain the desired mutation.
EN
Transformants containing the gene cassette and the desired mutations were selected, and by means of conventional crossing with wild-type plants, the CRISPR/Cas9 gene cassette was removed by selecting T1 plants that only contained the desired mutation, but no longer contained the gene cassette (null-segregants). Therefore, the plants in the field trial are not expected to contain foreign genetic material. They should only contain the desired mutation.
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BCH-GENE-SCBD-115122-1 NGAL2 transcription factor | Zea mays (Maize, Corn, MAIZE)Protein coding sequence | Changes in physiology and/or production (Yield)
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BCH-GENE-SCBD-107955-2 Plastochron 1 | Zea mays (Maize, Corn, MAIZE)Protein coding sequence | Changes in physiology and/or production (Growth rate, Reproduction, Yield),Tolerance to abiotic stress (Cold / Heat, Drought)
The insertion of a single basepair into the coding sequence of NGAL2 created a frameshift mutation that resulted in a knockoutof the gene. Thus, the native protein is not expected to be present in the modified cells. Moreover, since NGAL2 is a transcriptional repressor of PLA1, the absence of a functional NGAL2 should thus allow an increased expression of the PLA1 gene.
EN
- Research
EN
In Arabidopsis thaliana knockout experiments of NGAL2, seed size was enlarged, biomass was increased and inflorescent architecture was modified.
EN
- Addgene: pBUN411 [ English ]
- GMOInfo (JRC|EU): Notification B/BE/18/V8 [ English ]
- Belgian Biosafety server: B/BE/18/V8 [ English ]
