Specific targeting of point mutations in EGFR L858R-positive lung cancer by CRISPR/Cas9

Laboratory Investigation; a Journal of Technical Methods and Pathology
Alvin Ho-Kwan CheungKa-Fai To

Abstract

Cancer cells are defined genetically by the mutations they harbor, commonly single nucleotide substitutions. Therapeutic approaches which specifically target cancer cells by recognizing these defining genetic aberrations are expected to exhibit minimal side-effects. However, current protein-based targeted therapy is greatly limited by the range of genes that can be targeted, as well as by acquired resistance. We hypothesized that a therapeutic oligonucleotide-based strategy may address this need of specific cancer targeting. We used CRISPR/Cas9 system to target a commonly occurring EGFR point mutation, L858R, with an oligonucleotide guide that recognizes L858R as the suitable protospacer-adjacent motif (PAM) sequence for DNA cleavage. We found that this strategy, which utilized PAM to differentiate cancer mutation from normal, afforded high specificity to the extent of a single nucleotide substitution. The anti-L858R vehicle resulted in selective genome cleavage only in L858R mutant cells, as detected by Sanger sequencing and T7 Endonuclease I assay. Wild-type cells were unaffected by the same treatment. Digital PCR revealed 37.9 ± 8.57% of L858R gene copies were targeted in mutant. Only treated mutant cells, but not wild-type ...Continue Reading

References

Feb 24, 2007·Nature Reviews. Cancer·Sreenath V SharmaDaniel A Haber
Dec 21, 2007·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Gregory L Verdine, Loren D Walensky
Aug 21, 2009·The New England Journal of Medicine·Tony S MokMasahiro Fukuoka
Apr 7, 2011·Nature Reviews. Genetics·Mark A Kay
Apr 24, 2012·Genome Research·Eunji KimJin-Soo Kim
Jan 5, 2013·Science·Le CongFeng Zhang
Jan 5, 2013·Science·Prashant MaliGeorge M Church
Jan 26, 2013·Science·Franklin W HuangLevi A Garraway
Jun 25, 2013·Nature Biotechnology·Yanfang FuJeffry D Sander
Aug 16, 2013·Nature·Ludmil B AlexandrovMichael R Stratton
Sep 4, 2014·Nature Biotechnology·John G DoenchDavid E Root
Oct 18, 2014·Nature Reviews. Drug Discovery·Adrienne D CoxChanning J Der
Oct 20, 2014·Nature Biotechnology·Lukasz SwiechFeng Zhang
Oct 30, 2014·The New England Journal of Medicine·Donal P McLornanGhulam J Mufti
May 12, 2015·Nature Biotechnology·Junwei ShiChristopher R Vakoc
Jun 23, 2015·Nature·Benjamin P KleinstiverJ Keith Joung
Jan 20, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Ciaran M LeeGang Bao
Jan 28, 2016·Scientific Reports·Xiaoxi LiuToru Takumi
Nov 17, 2016·Nature·Keiichiro SuzukiJuan Carlos Izpisua Belmonte
Jan 5, 2017·The New England Journal of Medicine·Arthur A Levin

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Citations

Mar 22, 2020·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Hua Alexander HanBoon-Seng Soh
Jan 3, 2021·Molecular Biotechnology·Precilla S DaisyT S Anitha
Aug 20, 2019·Trends in Molecular Medicine·Chunyang JiangZhigang Zhao
May 18, 2020·Advanced Drug Delivery Reviews·Tao Wan, Yuan Ping
Jan 23, 2021·RSC Medicinal Chemistry·Xuan YangAndrey A Ivanov
Dec 20, 2020·International Journal of Molecular Sciences·Edyta JanikMichal Bijak
Apr 12, 2021·Biosensors & Bioelectronics·Chenqi NiuChong Zhang

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Methods Mentioned

BETA
transfection
PCR
electrophoresis
Assay
xenografts
deamination

Software Mentioned

RainDance Analyst
SPSS

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