000917380 001__ 917380
000917380 005__ 20230123101927.0
000917380 037__ $$aFZJ-2023-00594
000917380 041__ $$aEnglish
000917380 1001_ $$0P:(DE-HGF)0$$aPouramini, P$$b0$$eCorresponding author
000917380 1112_ $$aBotanik-Tagung International Conference of the German Society for Plant Sciences 2022$$cBonn$$d2023-08-28 - 2023-09-01$$wGermany
000917380 245__ $$aTargeted knockout of barley endosperm-specific storage proteins as a prerequisite for molecular farming purposes
000917380 260__ $$c2022
000917380 3367_ $$033$$2EndNote$$aConference Paper
000917380 3367_ $$2DataCite$$aOther
000917380 3367_ $$2BibTeX$$aINPROCEEDINGS
000917380 3367_ $$2DRIVER$$aconferenceObject
000917380 3367_ $$2ORCID$$aLECTURE_SPEECH
000917380 3367_ $$0PUB:(DE-HGF)6$$2PUB:(DE-HGF)$$aConference Presentation$$bconf$$mconf$$s1673846071_18271$$xAfter Call
000917380 520__ $$aThe concept for producing valuable proteins in plants is known as molecular farming. Besides transient expression using viral vector systems in tobacco leaves, the cereal grain is a natural bioreactor ideal for storing proteins under ambient conditions. Therefore, the cereal grain provides a cost-effective, easily scalable expression system for producing high-value proteins in the starchy endosperm. CRISPR-mediated targeted knockout of barley Hordein B1 family members was achieved, and Sanger and deep amplicon sequencing genotyped mutant plants. ELISA and Western blotting performed the abundance and quantification of the recombinant protein overexpressed in wildtype and mutant background. An automated phenotyping device evaluated the morphological changes of the mutant grains. To increase protein yield and overcome the competition between endogenous storage protein accumulation and the high-value protein, a targeted knockout of the Hordein B1 family members by using Cas9 technology was performed. The recombinant human Epidermal Groth Factor (EGF) was overexpressed in wildtype and horb1 mutant background as a proof-of-concept. Grains of the resulting horb1 mutants showed altered biometric traits, reduced total protein and hordein content, and delayed germination behaviour compared to wild type segregants. Western blotting and ELISA results confirmed a high EGF accumulation in the horb1 mutant background. In conclusion, the strategy of higher abundance of a recombinant protein in barley grains was successfully demonstrated by reducing endogenous storage protein accumulation. Further fine-tuning is required to minimize the germination disadvantage.
000917380 536__ $$0G:(DE-HGF)POF4-2171$$a2171 - Biological and environmental resources for sustainable use (POF4-217)$$cPOF4-217$$fPOF IV$$x0
000917380 7001_ $$0P:(DE-Juel1)165733$$aKoller, Robert$$b1$$ufzj
000917380 7001_ $$0P:(DE-Juel1)129333$$aHuber, Gregor$$b2$$ufzj
000917380 7001_ $$0P:(DE-Juel1)129315$$aFischbach, Andreas$$b3$$ufzj
000917380 7001_ $$0P:(DE-HGF)0$$aBirnisdóttir, Björg$$b4
000917380 7001_ $$0P:(DE-HGF)0$$aBjörnsson, JM$$b5
000917380 7001_ $$0P:(DE-HGF)0$$aHensel, G$$b6
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000917380 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)165733$$aForschungszentrum Jülich$$b1$$kFZJ
000917380 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)129333$$aForschungszentrum Jülich$$b2$$kFZJ
000917380 9101_ $$0I:(DE-588b)5008462-8$$6P:(DE-Juel1)129315$$aForschungszentrum Jülich$$b3$$kFZJ
000917380 9131_ $$0G:(DE-HGF)POF4-217$$1G:(DE-HGF)POF4-210$$2G:(DE-HGF)POF4-200$$3G:(DE-HGF)POF4$$4G:(DE-HGF)POF$$9G:(DE-HGF)POF4-2171$$aDE-HGF$$bForschungsbereich Erde und Umwelt$$lErde im Wandel – Unsere Zukunft nachhaltig gestalten$$vFür eine nachhaltige Bio-Ökonomie – von Ressourcen zu Produkten$$x0
000917380 9141_ $$y2022
000917380 9201_ $$0I:(DE-Juel1)IBG-2-20101118$$kIBG-2$$lPflanzenwissenschaften$$x0
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