Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis

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Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis. / Jin, Yu; Marquardt, Sebastian.

In: Bio-protocol, Vol. 10, No. 20, 3796, 2020.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Jin, Y & Marquardt, S 2020, 'Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis', Bio-protocol, vol. 10, no. 20, 3796. https://doi.org/10.21769/BioProtoc.3796

APA

Jin, Y., & Marquardt, S. (2020). Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis. Bio-protocol, 10(20), [3796]. https://doi.org/10.21769/BioProtoc.3796

Vancouver

Jin Y, Marquardt S. Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis. Bio-protocol. 2020;10(20). 3796. https://doi.org/10.21769/BioProtoc.3796

Author

Jin, Yu ; Marquardt, Sebastian. / Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis. In: Bio-protocol. 2020 ; Vol. 10, No. 20.

Bibtex

@article{fc95f6d8512e4ee28db2f356bc72e610,
title = "Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis",
abstract = "CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of larger regions generate loss-of-function alleles that offer a straightforward starting point for functional dissections of genomic loci. We present an easy-to-use strategy including a fast cloning dual-sgRNA vector linked to efficient isolation of heritable Cas9-free genomic deletions to rapidly and cost-effectively generate a targeted heritable genome deletion. This step-by-step protocol includes gRNA design, cloning strategy and mutation detection for Arabidopsis and may be adapted for other plant species.",
keywords = "CRISPR/Cas9, Cas9-free, dual-sgRNA, gBlock, Genomic deletion, FUNCTIONAL GENOMICS, GENE DELETION, CRISPR/CAS9, MUTATIONS, SYSTEM, DNA",
author = "Yu Jin and Sebastian Marquardt",
year = "2020",
doi = "10.21769/BioProtoc.3796",
language = "English",
volume = "10",
journal = "Bio-protocol",
issn = "2331-8325",
publisher = "bio-protocol",
number = "20",

}

RIS

TY - JOUR

T1 - Dual sgRNA-based Targeted Deletion of Large Genomic Regions and Isolation of Heritable Cas9-free Mutants in Arabidopsis

AU - Jin, Yu

AU - Marquardt, Sebastian

PY - 2020

Y1 - 2020

N2 - CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of larger regions generate loss-of-function alleles that offer a straightforward starting point for functional dissections of genomic loci. We present an easy-to-use strategy including a fast cloning dual-sgRNA vector linked to efficient isolation of heritable Cas9-free genomic deletions to rapidly and cost-effectively generate a targeted heritable genome deletion. This step-by-step protocol includes gRNA design, cloning strategy and mutation detection for Arabidopsis and may be adapted for other plant species.

AB - CRISPR/Cas9 system directed by a gene-specific single guide RNA (sgRNA) is an effective tool for genome editing such as deletions of few bases in coding genes. However, targeted deletion of larger regions generate loss-of-function alleles that offer a straightforward starting point for functional dissections of genomic loci. We present an easy-to-use strategy including a fast cloning dual-sgRNA vector linked to efficient isolation of heritable Cas9-free genomic deletions to rapidly and cost-effectively generate a targeted heritable genome deletion. This step-by-step protocol includes gRNA design, cloning strategy and mutation detection for Arabidopsis and may be adapted for other plant species.

KW - CRISPR/Cas9

KW - Cas9-free

KW - dual-sgRNA

KW - gBlock

KW - Genomic deletion

KW - FUNCTIONAL GENOMICS

KW - GENE DELETION

KW - CRISPR/CAS9

KW - MUTATIONS

KW - SYSTEM

KW - DNA

U2 - 10.21769/BioProtoc.3796

DO - 10.21769/BioProtoc.3796

M3 - Journal article

C2 - 33659450

VL - 10

JO - Bio-protocol

JF - Bio-protocol

SN - 2331-8325

IS - 20

M1 - 3796

ER -

ID: 251575278