About: Efficient TALEN construction for Bombyx mori gene targeting

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An Entity of Type : http://linked.opendata.cz/ontology/domain/vavai/Vysledek, within Data Space : linked.opendata.cz associated with source document(s)

Attributes	Values
rdf:type	skos:Concept http://linked.opendata.cz/ontology/domain/vavai/Vysledek
rdfs:seeAlso	http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0073458
Description	Engineered nucleases are artificial enzymes able to introduce double stranded breaks at desired genomic locations. The double stranded breaks start the error-prone repair process of non-homologous end-joining (NHEJ), which eventually leads to the induction of mutations at target sites. We showed earlier that ZFNs and TALENs are able to induce NHEJ mutations in the B. mori genome. In order to optimize our mutagenesis protocol, we modified one of the reported truncated TALEN scaffolds and optimized it for use in the B. mori embryo. We also established a novel B. mori somatic cell assay suitable for the preselection of highly efficient TALENs directly in the B. mori model system. We compared the efficiency of several TALEN pairs based on three different frameworks using the BmBLOS2 gene. The new active TALENs show one order of magnitude higher efficiency than those we used previously. We confirmed the utility of our improved protocol by mutagenesis of the autosomal gene, red egg (Bm-re) and showed that it allows obtaining homozygous mutants in G1. Our procedure minimizes the chance of failure in B. mori gene targeting experiments. Engineered nucleases are artificial enzymes able to introduce double stranded breaks at desired genomic locations. The double stranded breaks start the error-prone repair process of non-homologous end-joining (NHEJ), which eventually leads to the induction of mutations at target sites. We showed earlier that ZFNs and TALENs are able to induce NHEJ mutations in the B. mori genome. In order to optimize our mutagenesis protocol, we modified one of the reported truncated TALEN scaffolds and optimized it for use in the B. mori embryo. We also established a novel B. mori somatic cell assay suitable for the preselection of highly efficient TALENs directly in the B. mori model system. We compared the efficiency of several TALEN pairs based on three different frameworks using the BmBLOS2 gene. The new active TALENs show one order of magnitude higher efficiency than those we used previously. We confirmed the utility of our improved protocol by mutagenesis of the autosomal gene, red egg (Bm-re) and showed that it allows obtaining homozygous mutants in G1. Our procedure minimizes the chance of failure in B. mori gene targeting experiments. (en)
Title	Efficient TALEN construction for Bombyx mori gene targeting Efficient TALEN construction for Bombyx mori gene targeting (en)
skos:prefLabel	Efficient TALEN construction for Bombyx mori gene targeting Efficient TALEN construction for Bombyx mori gene targeting (en)
skos:notation	RIV/60077344:_____/13:00396418!RIV14-AV0-60077344
http://linked.open...avai/predkladatel	Biologické centrum AV ČR, v. v. i.
http://linked.open...avai/riv/aktivita	I P V
http://linked.open...avai/riv/aktivity	I, P(GAP305/10/2406), V
http://linked.open...iv/cisloPeriodika	9
http://linked.open...vai/riv/dodaniDat	2014
http://linked.open...aciTvurceVysledku	Žurovec, Michal Sajwan, Suresh
http://linked.open.../riv/druhVysledku	J - Článek v odborném periodiku
http://linked.open...iv/duvernostUdaju	S - Úplné a pravdivé údaje nepodléhající ochraně podle zvláštních právních předpisů
http://linked.open...titaPredkladatele	Biologické centrum AV ČR, v. v. i.
http://linked.open...dnocenehoVysledku	72093
http://linked.open...ai/riv/idVysledku	RIV/60077344:_____/13:00396418
http://linked.open...riv/jazykVysledku	eng - angličtina
http://linked.open.../riv/klicovaSlova	Bombyx mori (en)
http://linked.open.../riv/klicoveSlovo	Bombyx mori
http://linked.open...odStatuVydavatele	US - Spojené státy americké
http://linked.open...ontrolniKodProRIV	[BD653E263A7B]
http://linked.open...i/riv/nazevZdroje	PLoS ONE
http://linked.open...in/vavai/riv/obor	EB
http://linked.open...ichTvurcuVysledku	2 (xsd:int)
http://linked.open...cetTvurcuVysledku	8 (xsd:int)
http://linked.open...vavai/riv/projekt	Targeted mutagenesis in silkworm
http://linked.open...UplatneniVysledku	2013
http://linked.open...v/svazekPeriodika	8
http://linked.open...iv/tvurceVysledku	Žurovec, Michal Daimon, T. Osanai-Futahashi, M. Sajwan, Suresh Sezutsu, H. Takasu, Y. Tamura, T. Uchino, K.
http://linked.open...ain/vavai/riv/wos	000324695900013
issn	1932-6203
number of pages	11 (xsd:int)
http://bibframe.org/vocab/doi	10.1371/journal.pone.0073458

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