Skip to main content

Advertisement

SpringerLink
Log in

Application of CRISPR/Cas9 genome editing to the study and treatment of disease

  • Review Article
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

CRISPR/Cas is a microbial adaptive immune system that uses RNA-guided nucleases to cleave foreign genetic elements. The CRISPR/Cas9 method has been engineered from the type II prokaryotic CRISPR system and uses a single-guide RNA to target the Cas9 nuclease to a specific genomic sequence. Cas9 induces double-stranded DNA breaks which are repaired either by imperfect non-homologous end joining to generate insertions or deletions (indels) or, if a repair template is provided, by homology-directed repair. Due to its specificity, simplicity and versatility, the CRISPR/Cas9 system has recently emerged as a powerful tool for genome engineering in various species. This technology can be used to investigate the function of a gene of interest or to correct gene mutations in cells via genome editing, paving the way for future gene therapy approaches. Improvements to the efficiency of CRISPR repair, in particular to increase the rate of gene correction and to reduce undesired off-target effects, and the development of more effective delivery methods will be required for its broad therapeutic application.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Ain QU, Chung JY, Kim Y-H (2014) Current and future delivery systems for engineered nucleases: ZFN, TALEN and RGEN. J Control Release. doi:10.1016/j.jconrel.2014.12.036

  • Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero DA, Horvath P (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315:1709–1712

    Article  CAS  PubMed  Google Scholar 

  • Bolotin A, Quinquis B, Sorokin A, Ehrlich SD (2005) Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology 151:2551–2561

    Article  CAS  PubMed  Google Scholar 

  • Cho SW, Kim S, Kim JM, Kim J-S (2013) Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease. Nat Biotechnol 31:230–232

    Article  CAS  PubMed  Google Scholar 

  • Cho SW, Kim S, Kim Y, Kweon J, Kim HS, Bae S, Kim J-S (2014) Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases. Genome Res 24:132–141

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Choi PS, Meyerson M (2014) Targeted genomic rearrangements using CRISPR/Cas technology. Nat Commun 5:3728

    CAS  PubMed Central  PubMed  Google Scholar 

  • Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339:819–823

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Davis L, Maizels N (2014) Homology-directed repair of DNA nicks via pathways distinct from canonical double-strand break repair. Proc Natl Acad Sci 111:E924–E932

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Deltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao Y, Pirzada ZA, Eckert MR, Vogel J, Charpentier E (2011) CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature 471:602–607

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Dianov GL, Hübscher U (2013) Mammalian base excision repair: the forgotten archangel. Nucleic Acids Res 41:3483–3490

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K (2013) Enhanced efficiency of human pluripotent stem cell genome editing through replacing TALENs with CRISPRs. Cell Stem Cell 12:393–394

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Doudna JA, Charpentier E (2014) The new frontier of genome engineering with CRISPR–Cas9. Science. doi:10.1126/science.1258096

    PubMed Central  Google Scholar 

  • Folger KR, Wong EA, Wahl G, Capecchi MR (1982) Patterns of integration of DNA microinjected into cultured mammalian cells: evidence for homologous recombination between injected plasmid DNA molecules. Mol Cell Biol 2:1372–1387

    CAS  PubMed Central  PubMed  Google Scholar 

  • Friedland AE, Tzur YB, Esvelt KM, Colaiacovo MP, Church GM, Calarco JA (2013) Heritable genome editing in C. elegans via a CRISPR–Cas9 system. Nat Methods 10:741–743

    Article  CAS  PubMed  Google Scholar 

  • Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD (2013) High-frequency off-target mutagenesis induced by CRISPR–Cas nucleases in human cells. Nat Biotechnol 31:822–826

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Fu Y, Sander JD, Reyon D, Cascio VM, Joung JK (2014) Improving CRISPR–Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol 32:279–284

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gabriel R, Lombardo A, Arens A, Miller JC, Genovese P, Kaeppel C, Nowrouzi A, Bartholomae CC, Wang J, Friedman G, Holmes MC, Gregory PD, Glimm H, Schmidt M, Naldini L, von Kalle C (2011) An unbiased genome-wide analysis of zinc-finger nuclease specificity. Nat Biotechnol 29:816–823

    Article  CAS  PubMed  Google Scholar 

  • Gaj T, Guo J, Kato Y, Sirk SJ, Barbas CF (2012) Targeted gene knockout by direct delivery of zinc-finger nuclease proteins. Nat Methods 9:805–807

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gaj T, Gersbach CA, Barbas Iii CF (2013) ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Trends Biotechnol 31:397–405

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gasiunas G, Barrangou R, Horvath P, Siksnys V (2012) Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc Natl Acad Sci USA 109:E2579–E2586

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gratz SJ, Cummings AM, Nguyen JN, Hamm DC, Donohue LK, Harrison MM, Wildonger J, O’Connor-Giles KM (2013) Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics 194:1029–1035

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Güell M, Yang L, Church GM (2014) Genome editing assessment using CRISPR genome analyzer (CRISPR–GA). Bioinformatics 30:2968–2970

    Article  PubMed  Google Scholar 

  • Guilinger JP, Thompson DB, Liu DR (2014) Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nat Biotechnol 32:577–582

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gupta RM, Musunuru K (2014) Expanding the genetic editing tool kit: ZFNs, TALENs, and CRISPR–Cas9. J Clin Investig 124:4154–4161

    Article  CAS  PubMed  Google Scholar 

  • Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey ME, Thielke A, Aster JC, Regev A, Ebert BL (2014) Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR–Cas9 genome editing. Nat Biotechnol 32:941–946

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Horii T, Tamura D, Morita S, Kimura M, Hatada I (2013) Generation of an ICF syndrome model by efficient genome editing of human induced pluripotent stem cells using the CRISPR system. Int J Mol Sci 14:19774–19781

    Article  PubMed Central  PubMed  Google Scholar 

  • Hsu Patrick D, Lander Eric S, Zhang F (2014) Development and applications of CRISPR–Cas9 for genome engineering. Cell 157:1262–1278

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O, Cradick TJ, Marraffini LA, Bao G, Zhang F (2013) DNA targeting specificity of RNA-guided Cas9 nucleases. Nat Biotechnol 31:827–832

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson RT, Yeh JRJ, Joung JK (2013) Efficient genome editing in zebrafish using a CRISPR–Cas system. Nat Biotechnol 31:227–229

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A (1987) Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol 169:5429–5433

    CAS  PubMed Central  PubMed  Google Scholar 

  • Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA (2013) RNA-guided editing of bacterial genomes using CRISPR–Cas systems. Nat Biotechnol 31:233–239

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821

    Article  CAS  PubMed  Google Scholar 

  • Jinek M, Jiang F, Taylor DW, Sternberg SH, Kaya E, Ma E, Anders C, Hauer M, Zhou K, Lin S, Kaplan M, Iavarone AT, Charpentier E, Nogales E, Doudna JA (2014) Structures of Cas9 endonucleases reveal RNA-mediated conformational activation. Science. doi:10.1126/science.1247997

    PubMed Central  PubMed  Google Scholar 

  • Katic I, Großhans H (2013) Targeted heritable mutation and gene conversion by Cas9-CRISPR in caenorhabditis elegans. Genetics 195:1173–1176

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Kim S, Kim D, Cho SW, Kim J, Kim J-S (2014) Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Res 24:1012–1019

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Koike-Yusa H, Li Y, Tan EP, Velasco-Herrera MDC, Yusa K (2014) Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR–guide RNA library. Nat Biotechnol 32:267–273

    Article  CAS  PubMed  Google Scholar 

  • Li D, Qiu Z, Shao Y, Chen Y, Guan Y, Liu M, Li Y, Gao N, Wang L, Lu X, Zhao Y, Liu M (2013a) Heritable gene targeting in the mouse and rat using a CRISPR–Cas system. Nat Biotechnol 31:681–683

    Article  CAS  PubMed  Google Scholar 

  • Li W, Teng F, Li T, Zhou Q (2013b) Simultaneous generation and germline transmission of multiple gene mutations in rat using CRISPR–Cas systems. Nat Biotechnol 31:684–686

    Article  CAS  PubMed  Google Scholar 

  • Li HL, Fujimoto N, Sasakawa N, Shirai S, Ohkame T, Sakuma T, Tanaka M, Amano N, Watanabe A, Sakurai H, Yamamoto T, Yamanaka S, Hotta A (2015) Precise correction of the dystrophin gene in duchenne muscular dystrophy patient induced pluripotent stem cells by TALEN and CRISPR–Cas9. Stem Cell Rep 4:143–154

    Article  CAS  Google Scholar 

  • Lin Y, Cradick TJ, Brown MT, Deshmukh H, Ranjan P, Sarode N, Wile BM, Vertino PM, Stewart FJ, Bao G (2014) CRISPR/Cas9 systems have off-target activity with insertions or deletions between target DNA and guide RNA sequences. Nucleic Acids Res 42:7473–7485

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Long C, McAnally JR, Shelton JM, Mireault AA, Bassel-Duby R, Olson EN (2014) Prevention of muscular dystrophy in mice by CRISPR/Cas9-mediated editing of germline DNA. Science 345:1184–1188

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Maddalo D, Manchado E, Concepcion CP, Bonetti C, Vidigal JA, Han Y-C, Ogrodowski P, Crippa A, Rekhtman N, de Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system. Nature 516:423–427

    Article  CAS  PubMed  Google Scholar 

  • Maggio I, Holkers M, Liu J, Janssen JM, Chen X, Goncalves MAFV (2014) Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep. doi:10.1038/srep05105

    PubMed Central  PubMed  Google Scholar 

  • Makarova KS, Aravind L, Grishin NV, Rogozin IB, Koonin EV (2002) A DNA repair system specific for thermophilic Archaea and bacteria predicted by genomic context analysis. Nucleic Acids Res 30:482–496

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Makarova KS, Aravind L, Wolf Y, Koonin E (2011a) Unification of Cas protein families and a simple scenario for the origin and evolution of CRISPR–Cas systems. Biol Direct 6:38

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Makarova KS, Haft DH, Barrangou R, Brouns SJJ, Charpentier E, Horvath P, Moineau S, Mojica FJM, Wolf YI, Yakunin AF, van der Oost J, Koonin EV (2011b) Evolution and classification of the CRISPR–Cas systems. Nat Rev Microbiol 9:467–477

    Article  CAS  PubMed  Google Scholar 

  • Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang L, Church GM (2013a) CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat Biotechnol 31:833–838

    Article  CAS  PubMed  Google Scholar 

  • Mali P, Esvelt KM, Church GM (2013b) Cas9 as a versatile tool for engineering biology. Nat Methods 10:957–963

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM (2013c) RNA-guided human genome engineering via Cas9. Science 339:823–826

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Mladenov E, Iliakis G (2011) Induction and repair of DNA double strand breaks: the increasing spectrum of non-homologous end joining pathways. Mutat Res 711:61–72

    Article  CAS  PubMed  Google Scholar 

  • Montague TG, Cruz JM, Gagnon JA, Church GM, Valen E (2014) CHOPCHOP: a CRISPR/Cas9 and TALEN web tool for genome editing. Nucleic Acids Res 42:W401–W407

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O (2014) Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell 156:935–949

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Niu Y, Shen B, Cui Y, Chen Y, Wang J, Wang L, Kang Y, Zhao X, Si W, Li W, Xiang AP, Zhou J, Guo X, Bi Y, Si C, Hu B, Dong G, Wang H, Zhou Z, Li T, Tan T, Pu X, Wang F, Ji S, Zhou Q, Huang X, Ji W, Sha J (2014) Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos. Cell 156:836–843

    Article  CAS  PubMed  Google Scholar 

  • Pattanayak V, Lin S, Guilinger JP, Ma E, Doudna JA, Liu DR (2013) High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nat Biotechnol 31:839–843

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ramakrishna S, Kwaku Dad A-B, Beloor J, Gopalappa R, Lee S-K, Kim H (2014) Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA. Genome Res 24:1020–1027

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, Trevino A, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F (2013) Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell. doi:10.1016/j.cell.2013.08.021

    PubMed Central  PubMed  Google Scholar 

  • Sanchez-Rivera FJ, Papagiannakopoulos T, Romero R, Tammela T, Bauer MR, Bhutkar A, Joshi NS, Subbaraj L, Bronson RT, Xue W, Jacks T (2014) Rapid modelling of cooperating genetic events in cancer through somatic genome editing. Nature 516:428–431

    Article  CAS  PubMed  Google Scholar 

  • Sander JD, Joung JK (2014) CRISPR–Cas systems for editing, regulating and targeting genomes. Nat Biotechnol 32:347–355

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V (2011) The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res 39:9275–9282

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schwank G, Koo B-K, Sasselli V, Dekkers JF, Heo I, Demircan T, Sasaki N, Boymans S, Cuppen E, van der Ent CK, Nieuwenhuis EE, Beekman JM, Clevers H (2013) Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. Cell Stem Cell 13:653–658

    Article  CAS  PubMed  Google Scholar 

  • Semenova E, Jore MM, Datsenko KA, Semenova A, Westra ER, Wanner B, van der Oost J, Brouns SJJ, Severinov K (2011) Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence. Proc Natl Acad Sci 108:10098–10103

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F (2014) Genome-scale CRISPR–Cas9 knockout screening in human cells. Science 343:84–87

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Shen B, Zhang W, Zhang J, Zhou J, Wang J, Chen L, Wang L, Hodgkins A, Iyer V, Huang X, Skarnes WC (2014) Efficient genome modification by CRISPR–Cas9 nickase with minimal off-target effects. Nat Methods 11:399–402

    Article  CAS  PubMed  Google Scholar 

  • Smith C, Gore A, Yan W, Abalde-Atristain L, Li Z, He C, Wang Y, Brodsky RA, Zhang K, Cheng L, Ye Z (2014) Whole-genome sequencing analysis reveals high specificity of CRISPR/Cas9 and TALEN-based genome editing in human iPSCs. Cell Stem Cell 15:12–13

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Sternberg SH, Redding S, Jinek M, Greene EC, Doudna JA (2014) DNA interrogation by the CRISPR RNA-guided endonuclease Cas9. Nature 507:62–67

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Suzuki K, Yu C, Qu J, Li M, Yao X, Yuan T, Goebl A, Tang S, Ren R, Aizawa E, Zhang F, Xu X, Soligalla RD, Chen F, Kim J, Kim NY, Liao H-K, Benner C, Esteban CR, Jin Y, Liu G-H, Li Y, Izpisua Belmonte JC (2014) Targeted gene correction minimally impacts whole-genome mutational load in human-disease-specific induced pluripotent stem cell clones. Cell Stem Cell 15:31–36

    Article  CAS  PubMed  Google Scholar 

  • Torres R, Martin MC, Garcia A, Cigudosa JC, Ramirez JC, Rodriguez-Perales S (2014) Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system. Nat Commun 5:3964

    Article  CAS  PubMed  Google Scholar 

  • Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ, Aryee MJ, Joung JK (2014) Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat Biotechnol 32:569–576

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • van der Oost J, Westra ER, Jackson RN, Wiedenheft B (2014) Unravelling the structural and mechanistic basis of CRISPR–Cas systems. Nat Rev Microbiol 12:479–492

    Article  PubMed  Google Scholar 

  • Veres A, Gosis Bridget S, Ding Q, Collins R, Ragavendran A, Brand H, Erdin S, Cowan CA, Talkowski Michael E, Musunuru K (2014) Low incidence of off-target mutations in individual CRISPR–Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing. Cell Stem Cell 15:27–30

    Article  CAS  PubMed  Google Scholar 

  • Waaijers S, Portegijs V, Kerver J, Lemmens BBLG, Tijsterman M, van den Heuvel S, Boxem M (2013) CRISPR/Cas9-targeted mutagenesis in caenorhabditis elegans. Genetics 195:1187–1191

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang H, Yang H, Shivalila CS, Dawlaty MM, Cheng AW, Zhang F, Jaenisch R (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153:910–918

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wang T, Wei JJ, Sabatini DM, Lander ES (2014) Genetic screens in human cells using the CRISPR–Cas9 system. Science 343:80–84

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Wu Y, Liang D, Wang Y, Bai M, Tang W, Bao S, Yan Z, Li D, Li J (2013) Correction of a genetic disease in mouse via use of CRISPR–Cas9. Cell Stem Cell 13:659–662

    Article  CAS  PubMed  Google Scholar 

  • Wu Y, Zhou H, Fan X, Zhang Y, Zhang M, Wang Y, Xie Z, Bai M, Yin Q, Liang D, Tang W, Liao J, Zhou C, Liu W, Zhu P, Guo H, Pan H, Wu C, Shi H, Wu L, Tang F, Li J (2015) Correction of a genetic disease by CRISPR–Cas9-mediated gene editing in mouse spermatogonial stem cells. Cell Res 25:67–79

    Article  CAS  PubMed  Google Scholar 

  • Xiao A, Cheng Z, Kong L, Zhu Z, Lin S, Gao G, Zhang B (2014) CasOT: a genome-wide Cas9/gRNA off-target searching tool. Bioinformatics 30:1180–1182

    Article  CAS  Google Scholar 

  • Xie F, Ye L, Chang JC, Beyer AI, Wang J, Muench MO, Kan YW (2014a) Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac. Genome Res 24:1526–1533

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Xie S, Shen B, Zhang C, Huang X, Zhang Y (2014b) sgRNAcas9: a software package for designing CRISPR sgRNA and evaluating potential off-target cleavage sites. PLoS ONE 9:e100448

    Article  PubMed Central  PubMed  Google Scholar 

  • Yin H, Xue W, Chen S, Bogorad RL, Benedetti E, Grompe M, Koteliansky V, Sharp PA, Jacks T, Anderson DG (2014) Genome editing with Cas9 in adult mice corrects a disease mutation and phenotype. Nat Biotechnol 32:551–553

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yu Z, Ren M, Wang Z, Zhang B, Rong YS, Jiao R, Gao G (2013) Highly efficient genome modifications mediated by CRISPR/Cas9 in drosophila. Genetics 195:289–291

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Yui S, Nakamura T, Sato T, Nemoto Y, Mizutani T, Zheng X, Ichinose S, Nagaishi T, Okamoto R, Tsuchiya K, Clevers H, Watanabe M (2012) Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+ stem cell. Nat Med 18:618–623

    Article  CAS  PubMed  Google Scholar 

  • Zhang F, Wen Y, Guo X (2014) CRISPR/Cas9 for genome editing: progress, implications and challenges. Hum Mol Genet 23:R40–R46

    Article  CAS  PubMed  Google Scholar 

  • Zhou J, Shen B, Zhang W, Wang J, Yang J, Chen L, Zhang N, Zhu K, Xu J, Hu B, Leng Q, Huang X (2014a) One-step generation of different immunodeficient mice with multiple gene modifications by CRISPR/Cas9 mediated genome engineering. Int J Biochem Cell Biol 46:49–55

    Article  CAS  PubMed  Google Scholar 

  • Zhou Y, Zhu S, Cai C, Yuan P, Li C, Huang Y, Wei W (2014b) High-throughput screening of a CRISPR/Cas9 library for functional genomics in human cells. Nature 509:487–491

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

AP and JB acknowledge the support of Leukaemia and Lymphoma Research (UK).

Conflict of interest

The authors declare that there are no conflicts of interest.

Author information

Authors and Affiliations

  1. Leukaemia and Lymphoma Research Molecular Haematology Unit, Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, and NIHR Biomedical Research Centre, Oxford University Hospitals, University of Oxford, Oxford, OX3 9DU, UK

    Andrea Pellagatti, Hamid Dolatshad, Simona Valletta & Jacqueline Boultwood

Authors
  1. Andrea Pellagatti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hamid Dolatshad
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simona Valletta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jacqueline Boultwood
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jacqueline Boultwood.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Pellagatti, A., Dolatshad, H., Valletta, S. et al. Application of CRISPR/Cas9 genome editing to the study and treatment of disease. Arch Toxicol 89, 1023–1034 (2015). https://doi.org/10.1007/s00204-015-1504-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-015-1504-y

Keywords

Search

Navigation