Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system
Phalaenopsis amabilis (L.) Blume commonly called Moth Orchid (Orchidaceae) is a natural orchid species designated as the National Flower of Indonesia for its beautiful flower shape and long-lasting flowering period. Basically, P. amabilis has a long vegetative phase that cause late flowering, about...
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2024
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| author | Suputri, N.P.A.E.O. Prasojo, I.S. Prabowo, L.A.T. Purwestri, Y.A. Purnomo, Purnomo Semiarti, E. |
| author_facet | Suputri, N.P.A.E.O. Prasojo, I.S. Prabowo, L.A.T. Purwestri, Y.A. Purnomo, Purnomo Semiarti, E. |
| author_sort | Suputri, N.P.A.E.O. |
| collection | UGM |
| description | Phalaenopsis amabilis (L.) Blume commonly called Moth Orchid (Orchidaceae) is a natural orchid species designated as the National Flower of Indonesia for its beautiful flower shape and long-lasting flowering period. Basically, P. amabilis has a long vegetative phase that cause late flowering, about 2 to 3 years for flowering, hence a method to shorten vegetative period is desired. The latest technological approach that can be used to accelerate flowering of P. amabilis is the CRISPR/Cas9 genome editing method to inactivate the GAI (Gibberellic Acid Insensitive) gene as a mutant gene that can accelerate the regulation of FLOWERING TIME (FT) genes flowering biosynthesis pathway. The approach that needs to be taken is to silence the GAI gene with a knockout system which begins with identifying and characterizing the GAI target gene in the P. amabilis which will be used as a single guide RNA. CRISPR/Cas9 mediated knockout efficiency is highly dependent on the properties of the sgRNA used. SgRNA consists of a target sequence, determining its specificity performance. We executed phylogenetic clustering for the PaGAI protein with closely related orchid species such as Dendrobium capra, Dendrobium cultivars and Cymbidium sinensis. SWISS-Model as tool webserver for protein structure homology modeling. Results show that P. amabilis has a specific domain with the occurrence of point mutations in the two conservative domains. Therefore, a single guide RNA reconstruction needs to be implemented. © 2024, Instituto Internacional de Ecologia. All rights reserved. |
| first_indexed | 2024-03-14T00:03:04Z |
| format | Article |
| id | oai:generic.eprints.org:279106 |
| institution | Universiti Gadjah Mada |
| last_indexed | 2024-03-14T00:03:04Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | oai:generic.eprints.org:2791062023-11-02T01:08:45Z https://repository.ugm.ac.id/279106/ Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system Suputri, N.P.A.E.O. Prasojo, I.S. Prabowo, L.A.T. Purwestri, Y.A. Purnomo, Purnomo Semiarti, E. Molecular Evolution Plant Biology Biological Sciences Phalaenopsis amabilis (L.) Blume commonly called Moth Orchid (Orchidaceae) is a natural orchid species designated as the National Flower of Indonesia for its beautiful flower shape and long-lasting flowering period. Basically, P. amabilis has a long vegetative phase that cause late flowering, about 2 to 3 years for flowering, hence a method to shorten vegetative period is desired. The latest technological approach that can be used to accelerate flowering of P. amabilis is the CRISPR/Cas9 genome editing method to inactivate the GAI (Gibberellic Acid Insensitive) gene as a mutant gene that can accelerate the regulation of FLOWERING TIME (FT) genes flowering biosynthesis pathway. The approach that needs to be taken is to silence the GAI gene with a knockout system which begins with identifying and characterizing the GAI target gene in the P. amabilis which will be used as a single guide RNA. CRISPR/Cas9 mediated knockout efficiency is highly dependent on the properties of the sgRNA used. SgRNA consists of a target sequence, determining its specificity performance. We executed phylogenetic clustering for the PaGAI protein with closely related orchid species such as Dendrobium capra, Dendrobium cultivars and Cymbidium sinensis. SWISS-Model as tool webserver for protein structure homology modeling. Results show that P. amabilis has a specific domain with the occurrence of point mutations in the two conservative domains. Therefore, a single guide RNA reconstruction needs to be implemented. © 2024, Instituto Internacional de Ecologia. All rights reserved. 2024 Article PeerReviewed Suputri, N.P.A.E.O. and Prasojo, I.S. and Prabowo, L.A.T. and Purwestri, Y.A. and Purnomo, Purnomo and Semiarti, E. (2024) Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system. Brazilian Journal of Biology, 84. https://www.scopus.com/inward/record.uri?eid=2-s2.0-85161166984&doi=10.1590%2f1519-6984.268133&partnerID=40&md5=6a26dc134c63748e6d20a6063a93fb79 |
| spellingShingle | Molecular Evolution Plant Biology Biological Sciences Suputri, N.P.A.E.O. Prasojo, I.S. Prabowo, L.A.T. Purwestri, Y.A. Purnomo, Purnomo Semiarti, E. Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title | Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title_full | Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title_fullStr | Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title_full_unstemmed | Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title_short | Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system |
| title_sort | identification of early flowering mutant gene in phalaenopsis amabilis l blume for sgrna construction in crispr cas9 genome editing system |
| topic | Molecular Evolution Plant Biology Biological Sciences |
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