Institut for Biomedicin

Lars Bolund

Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes

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Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes. / Vad-Nielsen, Johan; Lin, Lin; Bolund, Lars; Nielsen, Anders Lade; Luo, Yonglun.

I: Cellular and Molecular Life Sciences, Bind 73, Nr. 22, 13.05.2016, s. 4315–4325.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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@article{eae7104f99804fdaa3163e96efaf7e70,
title = "Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes",
abstract = "The engineered CRISPR/Cas9 technology has developed as the most efficient and broadly used genome editing tool. However, simultaneously targeting multiple genes (or genomic loci) in the same individual cells using CRISPR/Cas9 remain one technical challenge. In this article, we have developed a Golden Gate Assembly method for the generation of CRISPR gRNA expression arrays, thus enabling simultaneous gene targeting. Using this method, the generation of CRISPR gRNA expression array can be accomplished in 2 weeks, and contains up to 30 gRNA expression cassettes. We demonstrated in the study that simultaneously targeting 10 genomic loci or simultaneously inhibition of multiple endogenous genes could be achieved using the multiplexed gRNA expression array vector in human cells. The complete set of plasmids is available through the non-profit plasmid repository Addgene.",
author = "Johan Vad-Nielsen and Lin Lin and Lars Bolund and Nielsen, {Anders Lade} and Yonglun Luo",
year = "2016",
month = may,
day = "13",
doi = "10.1007/s00018-016-2271-5",
language = "English",
volume = "73",
pages = "4315–4325",
journal = "Cellular and Molecular Life Sciences",
issn = "1420-682X",
publisher = "Springer Basel AG",
number = "22",

}

RIS

TY - JOUR

T1 - Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes

AU - Vad-Nielsen, Johan

AU - Lin, Lin

AU - Bolund, Lars

AU - Nielsen, Anders Lade

AU - Luo, Yonglun

PY - 2016/5/13

Y1 - 2016/5/13

N2 - The engineered CRISPR/Cas9 technology has developed as the most efficient and broadly used genome editing tool. However, simultaneously targeting multiple genes (or genomic loci) in the same individual cells using CRISPR/Cas9 remain one technical challenge. In this article, we have developed a Golden Gate Assembly method for the generation of CRISPR gRNA expression arrays, thus enabling simultaneous gene targeting. Using this method, the generation of CRISPR gRNA expression array can be accomplished in 2 weeks, and contains up to 30 gRNA expression cassettes. We demonstrated in the study that simultaneously targeting 10 genomic loci or simultaneously inhibition of multiple endogenous genes could be achieved using the multiplexed gRNA expression array vector in human cells. The complete set of plasmids is available through the non-profit plasmid repository Addgene.

AB - The engineered CRISPR/Cas9 technology has developed as the most efficient and broadly used genome editing tool. However, simultaneously targeting multiple genes (or genomic loci) in the same individual cells using CRISPR/Cas9 remain one technical challenge. In this article, we have developed a Golden Gate Assembly method for the generation of CRISPR gRNA expression arrays, thus enabling simultaneous gene targeting. Using this method, the generation of CRISPR gRNA expression array can be accomplished in 2 weeks, and contains up to 30 gRNA expression cassettes. We demonstrated in the study that simultaneously targeting 10 genomic loci or simultaneously inhibition of multiple endogenous genes could be achieved using the multiplexed gRNA expression array vector in human cells. The complete set of plasmids is available through the non-profit plasmid repository Addgene.

U2 - 10.1007/s00018-016-2271-5

DO - 10.1007/s00018-016-2271-5

M3 - Journal article

C2 - 27178736

VL - 73

SP - 4315

EP - 4325

JO - Cellular and Molecular Life Sciences

JF - Cellular and Molecular Life Sciences

SN - 1420-682X

IS - 22

ER -