TY  - GEN
A1  - Lukan, Tjaša
A1  - Machens, Fabian
A1  - Coll, Anna
A1  - Baebler, Špela
A1  - Messerschmidt, Katrin
A1  - Gruden, Kristina
T1  - Plant X-tender
BT  - an extension of the AssemblX system for the assembly and expression of multigene constructs in plants
T2  - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
N2  - Cloning multiple DNA fragments for delivery of several genes of interest into the plant genome is one of the main technological challenges in plant synthetic biology. Despite several modular assembly methods developed in recent years, the plant biotechnology community has not widely adopted them yet, probably due to the lack of appropriate vectors and software tools. Here we present Plant X-tender, an extension of the highly efficient, scarfree and sequence-independent multigene assembly strategy AssemblX,based on overlapdepended cloning methods and rare-cutting restriction enzymes. Plant X-tender consists of a set of plant expression vectors and the protocols for most efficient cloning into the novel vector set needed for plant expression and thus introduces advantages of AssemblX into plant synthetic biology. The novel vector set covers different backbones and selection markers to allow full design flexibility. We have included ccdB counterselection, thereby allowing the transfer of multigene constructs into the novel vector set in a straightforward and highly efficient way. Vectors are available as empty backbones and are fully flexible regarding the orientation of expression cassettes and addition of linkers between them, if required. We optimised the assembly and subcloning protocol by testing different scar-less assembly approaches: the noncommercial SLiCE and TAR methods and the commercial Gibson assembly and NEBuilder HiFi DNA assembly kits. Plant X-tender was applicable even in combination with low efficient homemade chemically competent or electrocompetent Escherichia coli. We have further validated the developed procedure for plant protein expression by cloning two cassettes into the newly developed vectors and subsequently transferred them to Nicotiana benthamiana in a transient expression setup. Thereby we show that multigene constructs can be delivered into plant cells in a streamlined and highly efficient way. Our results will support faster introduction of synthetic biology into plant science.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 990 
KW  - ligation cloning extract
KW  - DNA cloning
KW  - synthetic biology
KW  - multiple genes
KW  - vector system
KW  - transformation
KW  - recombination
KW  - protein
KW  - RNA
KW  - Methylation
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-446281
SN  - 1866-8372
IS  - 990
ER  - 
TY  - JOUR
A1  - Ran, Niva A.
A1  - Love, John A.
A1  - Heiber, Michael C.
A1  - Jiao, Xuechen
A1  - Hughes, Michael P.
A1  - Karki, Akchheta
A1  - Wang, Ming
A1  - Brus, Viktor V.
A1  - Wang, Hengbin
A1  - Neher, Dieter
A1  - Ade, Harald
A1  - Bazan, Guillermo C.
A1  - Thuc-Quyen Nguyen, 
T1  - Charge generation and recombination in an organic solar cell with low energetic offsets
JF  - dvanced energy materials
N2  - Organic bulk heterojunction (BHJ) solar cells require energetic offsets between the donor and acceptor to obtain high short-circuit currents (J(SC)) and fill factors (FF). However, it is necessary to reduce the energetic offsets to achieve high open-circuit voltages (V-OC). Recently, reports have highlighted BHJ blends that are pushing at the accepted limits of energetic offsets necessary for high efficiency. Unfortunately, most of these BHJs have modest FF values. How the energetic offset impacts the solar cell characteristics thus remains poorly understood. Here, a comprehensive characterization of the losses in a polymer:fullerene BHJ blend, PIPCP:phenyl-C61-butyric acid methyl ester (PC61BM), that achieves a high V-OC (0.9 V) with very low energy losses (E-loss = 0.52 eV) from the energy of absorbed photons, a respectable J(SC) (13 mA cm(-2)), but a limited FF (54%) is reported. Despite the low energetic offset, the system does not suffer from field-dependent generation and instead it is characterized by very fast nongeminate recombination and the presence of shallow traps. The charge-carrier losses are attributed to suboptimal morphology due to high miscibility between PIPCP and PC61BM. These results hold promise that given the appropriate morphology, the J(SC), V-OC, and FF can all be improved, even with very low energetic offsets.
KW  - energetic offset
KW  - fill factor
KW  - morphology
KW  - organic solar cells
KW  - recombination
Y1  - 2018
U6  - https://doi.org/10.1002/aenm.201701073
SN  - 1614-6832
SN  - 1614-6840
VL  - 8
IS  - 5
PB  - Wiley-VCH
CY  - Weinheim
ER  -