High efficient multisites genome editing in allotetraploid cotton (Gossypium hirsutum) using CRISPR/Cas9 system

Plant Biotechnology Journal
Pengcheng WangXianlong Zhang

Abstract

Gossypium hirsutum is an allotetraploid with a complex genome. Most genes have multiple copies that belong to At and Dt subgenomes. Sequence similarity is also very high between gene homologues. To efficiently achieve site/gene-specific mutation is quite needed. Due to its high efficiency and robustness, the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 system has exerted broad site-specific genome editing from prokaryotes to eukaryotes. In this study, we utilized a CRISPR/Cas9 system to generate two sgRNAs in a single vector to conduct multiple sites genome editing in allotetraploid cotton. An exogenously transformed gene Discosoma red fluorescent protein2(DsRed2) and an endogenous gene GhCLA1 were chosen as targets. The DsRed2-edited plants in T0 generation reverted its traits to wild type, with vanished red fluorescence the whole plants. Besides, the mutated phenotype and genotype were inherited to their T1 progenies. For the endogenous gene GhCLA1, 75% of regenerated plants exhibited albino phenotype with obvious nucleotides and DNA fragments deletion. The efficiency of gene editing at each target site is 66.7-100%. The mutation genotype was checked for both genes with Sanger sequencing. Barcode-ba...Continue Reading

References

May 1, 1996·The Plant Journal : for Cell and Molecular Biology·M A MandelP León
Dec 12, 2001·The Plant Journal : for Cell and Molecular Biology·G JachJ Schell
Jan 9, 2010·Science·Philippe Horvath, Rodolphe Barrangou
Apr 2, 2011·Nature·Elitza DeltchevaEmmanuelle Charpentier
Sep 14, 2011·Annual Review of Genetics·Lorraine S Symington, Jean Gautier
Nov 9, 2011·Annual Review of Genetics·Devaki BhayaRodolphe Barrangou
Jun 30, 2012·Science·Martin JinekEmmanuelle Charpentier
Jan 5, 2013·Science·Le CongFeng Zhang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Jan 31, 2013·Nature Biotechnology·Woong Y HwangJ Keith Joung
May 15, 2013·Trends in Biotechnology·Thomas GajCarlos F Barbas
Jun 25, 2013·Nature Biotechnology·Yanfang FuJeffry D Sander
Jul 23, 2013·Nature Biotechnology·Patrick D HsuFeng Zhang
Aug 10, 2013·Nature Biotechnology·Qiwei ShanCaixia Gao
Aug 10, 2013·Nature Biotechnology·Vladimir NekrasovSophien Kamoun
Aug 21, 2013·Molecular Plant·Kabin Xie, Yinong Yang
Aug 21, 2013·Cell Research·Zhengyan FengJian-Kang Zhu
Sep 4, 2013·Cell Research·Jin MiaoLi-Jia Qu
Oct 26, 2013·Nature Protocols·F Ann RanFeng Zhang
Dec 18, 2013·Science·Tim WangEric S Lander
Mar 4, 2014·Nature Biotechnology·Jeffry D Sander, J Keith Joung
May 20, 2014·Nature Genetics·Fuguang LiShuxun Yu
Sep 19, 2014·Nature Protocols·Qiwei ShanCaixia Gao
Oct 14, 2014·Cell·Luke A GilbertJonathan S Weissman
Mar 4, 2015·Proceedings of the National Academy of Sciences of the United States of America·Kabin XieYinong Yang
Apr 14, 2015·Nature Biotechnology·Rahul SatijaAviv Regev
Apr 17, 2015·BMC Biotechnology·Thomas B JacobsWayne A Parrott
May 30, 2015·Scientific Reports·Xianjun SunYajun Xi

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Citations

Oct 3, 2018·International Journal of Molecular Sciences·Shouhong ZhuJie Sun
Nov 6, 2018·Journal of Integrative Plant Biology·Jochen KumlehnHolger Puchta
Jan 17, 2019·Critical Reviews in Biotechnology·Aili BaoLam-Son Phan Tran
Apr 20, 2019·Plant Biotechnology Journal·Yiwang ZhuFeng Wang
Jul 19, 2019·Journal of Cellular Physiology·Niaz AhmadBaohong Zhang
Dec 29, 2019·Plant Biotechnology Journal·Wei GaoChun-Peng Song
Nov 7, 2019·Journal of Integrative Plant Biology·Sylvans OcholaSuomeng Dong
Dec 19, 2017·Plant Biotechnology Journal·Yang YangYongming Zhou
Aug 28, 2019·Plant Biotechnology Journal·Mengyan BaiYuefeng Guan
May 28, 2019·Molecular Genetics and Genomics : MGG·Xingpeng XiongJiashu Cao
Aug 22, 2017·Frontiers in Plant Science·Wei GaoChunpeng Song
Oct 6, 2018·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Wei ChenYongshan Zhang
Jun 6, 2020·Frontiers in Plant Science·Marcos Fernando BassoMaria Fatima Grossi-de-Sa
Apr 5, 2019·Plant Biotechnology Journal·Marcos Fernando BassoMaria Fatima Grossi-de-Sa
Nov 10, 2017·Emerging Topics in Life Sciences·Jorge Martínez-FortúnHuw D Jones

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Methods Mentioned

BETA
transgenic
phosphotransferase
gene knockout
PCR
Genotyping
gene knockdown
electrophoresis

Software Mentioned

Perl script
Trimmomatic
BioEdit
ot2gtf _

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