A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F17%3A00485647" target="_blank" >RIV/68081707:_____/17:00485647 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1105/tpc.16.00922" target="_blank" >http://dx.doi.org/10.1105/tpc.16.00922</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1105/tpc.16.00922" target="_blank" >10.1105/tpc.16.00922</a>

Result language

angličtina

Original language name

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

Original language description

We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination. Regulation of transcription is also possible. A Web-based tool streamlines vector selection and construction. One advantage of our platform is the use of the Csy-type (CRISPR system yersinia) ribonuclease 4 (Csy4) and tRNA processing enzymes to simultaneously express multiple guide RNAs (gRNAs). For example, we demonstrate targeted deletions in up to six genes by expressing 12 gRNAs from a single transcript. Csy4 and tRNA expression systems are almost twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promoters. Mutagenesis can be further enhanced 2.5-fold by incorporating the Trex2 exonuclease. Finally, we demonstrate that Cas9 nickases induce gene targeting at frequencies comparable to native Cas9 when they are delivered on geminivirus replicons. The reagents have been successfully validated in tomato (Solanum lycopersicum), tobacco (Nicotiana tabacum), Medicago truncatula, wheat (Triticum aestivum), and barley (Hordeum vulgare).

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

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Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Plant Cell

ISSN

1040-4651

e-ISSN

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Volume of the periodical

29

Issue of the periodical within the volume

6

Country of publishing house

US - UNITED STATES

Number of pages

22

Pages from-to

1196-1217

UT code for WoS article

000404989600006

EID of the result in the Scopus database

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