Augmenting CRISPR applications in Drosophila with tRNA-flanked sgRNAs

被引：289

作者：

Port, Fillip ^{[1
,2
]}

Bullock, Simon L. ^{[1
]}

机构：

[1] MRC Lab Mol Biol, Div Cell Biol, Cambridge, England

[2] German Canc Res Ctr, Div Signaling & Funct Genom, Heidelberg, Germany

来源：

NATURE METHODS | 2016年 / 13卷 / 10期

关键词：

HUMAN-CELLS; GENE DRIVES; IN-VIVO; CPF1; ACTIVATION; NUCLEASES; CAS9; SPECIFICITIES; MUTAGENESIS; SYSTEM;

D O I：

10.1038/nmeth.3972

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We present tRNA-based vectors for producing multiple clustered regularly interspaced short palindromic repeats (CRISPR) single guide RNAs (sgRNAs) from a single RNA polymerase II or III transcript in Drosophila. The system, which is based on liberation of sgRNAs by processing flanking tRNAs, permits highly efficient multiplexing of Cas9-based mutagenesis. We also demonstrate that the tRNA-sgRNA system markedly increases the efficacy of conditional gene disruption by Cas9 and can promote editing by the recently discovered RNA-guided endonuclease Cpf1.

引用

页码：852 / +

页数：5

共 26 条

[1] Cheating evolution: engineering gene drives to manipulate the fate of wild populations [J].

Champer, Jackson ;

Buchman, Anna ;

Akbari, Omar S. .

NATURE REVIEWS GENETICS, 2016, 17 (03) :146-159

[2] Optimizing sgRNA structure to improve CRISPR-Cas9 knockout efficiency [J].

Dang, Ying ;

Jia, Gengxiang ;

Choi, Jennie ;

Ma, Hongming ;

Anaya, Edgar ;

Ye, Chunting ;

Shankar, Premlata ;

Wu, Haoquan .

GENOME BIOLOGY, 2015, 16

[3] The new frontier of genome engineering with CRISPR-Cas9 [J].

Doudna, Jennifer A. ;

Charpentier, Emmanuelle .

SCIENCE, 2014, 346 (6213) :1077-+

[4] Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins [J].

Hur, Junho K. ;

Kim, Kyoungmi ;

Been, Kyung Wook ;

Baek, Gayoung ;

Ye, Sunghyeok ;

Hur, Junseok W. ;

Ryu, Seuk-Min ;

Lee, Youn Su ;

Kim, Jin-Soo .

NATURE BIOTECHNOLOGY, 2016, 34 (08) :807-808

[5]

Kim D, 2016, NAT BIOTECHNOL, V34, P863, DOI 10.1038/nbt.3609

[6] Generation of knockout mice by Cpf1-mediated gene targeting [J].

Kim, Yongsub ;

Cheong, Seung-A ;

Lee, Jong Geol ;

Lee, Sang-Wook ;

Lee, Myeong Sup ;

Baek, In-Jeoung ;

Sung, Young Hoon .

NATURE BIOTECHNOLOGY, 2016, 34 (08) :808-810

[7] Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells [J].

Kleinstiver, Benjamin P. ;

Tsai, Shengdar Q. ;

Prew, Michelle S. ;

Nguyen, Nhu T. ;

Welch, Moira M. ;

Lopez, Jose M. ;

McCaw, Zachary R. ;

Aryee, Martin J. ;

Joung, J. Keith .

NATURE BIOTECHNOLOGY, 2016, 34 (08) :869-+

[8] High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects [J].

Kleinstiver, Benjamin P. ;

Pattanayak, Vikram ;

Prew, Michelle S. ;

Tsai, Shengdar Q. ;

Nguyen, Nhu T. ;

Zheng, Zongli ;

Joung, J. Keith .

NATURE, 2016, 529 (7587) :490-+

[9] In Vivo Transcriptional Activation Using CRISPR/Cas9 in Drosophila [J].

Lin, Shuailiang ;

Ewen-Campen, Ben ;

Ni, Xiaochun ;

Housden, Benjamin E. ;

Perrimon, Norbert .

GENETICS, 2015, 201 (02) :433-+

[10]

Maeder ML, 2013, NAT METHODS, V10, P977, DOI [10.1038/nmeth.2598, 10.1038/NMETH.2598]

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