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Rebooting Quinoa – CRISPR-Combo Edition

Beschreibung

Genetic transformation of quinoa remains a major challenge. A stable transformation method has been developed for the accession QQ74, but it requires a laborious in vitro floral culture system and several months of selection (https://doi.org/10.1007/s11627-024-10450-z). More recently, Xie et al. published a highly efficient regeneration protocol based on hypocotyl-derived callus from the quinoa cultivar Qingqua I (https://doi.org/10.1038/s41598-025-03598-0). However, when we tested this protocol in QQ74, the tissues produced callus but failed to regenerate shoots.

What if this failure could become the perfect experimental system?

Because the Xie protocol does not regenerate QQ74 on its own, it provides a clear baseline for testing whether the activation of morphogenic genes can rescue regeneration. This is where CRISPR-Combo comes into play :

gene editing ← CRISPR-Combo → gene activation

CRISPR-Combo uses the same Cas9-based system for two different purposes: conventional guide RNAs direct genome editing, while specialized truncated guide RNAs recruit transcriptional activators to switch on endogenous genes. Ramasamy et al. recently demonstrated that activating morphogenic genes with this system can improve or accelerate regeneration in several recalcitrant plant species (https://doi.org/10.1038/s41467-026-76367-w).

In this project, we will first establish an Agrobacterium tumefaciens-mediated transformation workflow for QQ74 using strain GV3101 carrying pCAMBIA2301. GUS activity and paromomycin resistance will be used to identify transformed tissues and optimize the transformation conditions.

We will then introduce CRISPR-Combo constructs into the non-regenerating QQ74 tissue-culture system. By comparing CRISPR-Combo-treated explants with the corresponding controls, we will test whether activating endogenous morphogenic genes (BBM, WUS, etc) can restart shoot formation where the regeneration protocol alone fails.

In this project, you will learn to:

• Work with quinoa tissue culture and callus induction
• Perform Agrobacterium-mediated genetic transformation
• Use GUS assays and paromomycin selection to identify transformed tissues
• Distinguish transformation efficiency from regeneration efficiency
• Design guide RNAs for genome editing and gene activation
• Perform cloning
• Test whether morphogenic gene activation can rescue plant regeneration

Quinoa is difficult to transform—but perhaps the right genes simply need to be switched on.

Are you ready to reboot quinoa with CRISPR-Combo?

Projektzeitraum

Wintersemester 2026/2027

Bewerbungszeitraum

12. bis 26.10.2026

Durchführung

semesterbegleitend

Details zu Projektzeitraum und Durchführung

Flexible project plan and scope of work to meet your specific requirements.

Studienfach

Agrarbiologie
Agrarwissenschaften
Biologie
Biologie - Lehramt

Betreuende

Dr. Romano de Jesus Porras Murillo

Institut

Institut für Kulturpflanzenwissenschaften (340) (Physiology of Yield Stability (340k))

Sprache

deutsch/englisch

Teilnehmendenanzahl

min. 1, max. 1

Arbeitsaufwand

ca. 90 Stunden pro Teilnehmende:r | 3 ECTS-Punkte

Arbeitsaufwand (Stunden und ggf. ECTS) sind ungefähre Angaben. Die tatsächlich vergebenen ECTS-Punkte ergeben sich aus der tatsächlich geleisteten Arbeit.

 

Für dieses Projekt ist kein Motivationsschreiben des Studierenden erforderlich

Projektart

experimentell

Lernziele

Die Teilnehmende lernen in diesem Projekt:

• Work with quinoa tissue culture and callus induction
• Perform Agrobacterium-mediated genetic transformation
• Use GUS assays and paromomycin selection to identify transformed tissues
• Distinguish transformation efficiency from regeneration efficiency
• Design guide RNAs for genome editing and gene activation
• Perform cloning
• Test whether morphogenic gene activation can rescue plant regeneration

Anmerkungen für Studierende

Schlagworte

Gentechnik, Pflanzen, Quinoa, CRISPR, tissue culture, gene activation