{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T06:10:18Z","timestamp":1783750218280,"version":"3.55.0"},"reference-count":88,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2019,1,21]],"date-time":"2019-01-21T00:00:00Z","timestamp":1548028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000861","name":"Burroughs Wellcome Fund","doi-asserted-by":"publisher","award":["PDEP"],"award-info":[{"award-number":["PDEP"]}],"id":[{"id":"10.13039\/100000861","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Brain Sciences"],"abstract":"<jats:p>Gene-editing using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) is promising as a potential therapeutic strategy for many genetic disorders. CRISPR-based therapies are already being assessed in clinical trials, and evaluation of this technology in Fragile X syndrome has been performed by a number of groups. The findings from these studies and the advancement of CRISPR-based technologies are insightful as the field continues towards treatments and cures of Fragile X-Associated Disorders (FXADs). In this review, we summarize reports using CRISPR-editing strategies to target Fragile X syndrome (FXS) molecular dysregulation, and highlight how differences in FXS and Fragile X-associated Tremor\/Ataxia Syndrome (FXTAS) might alter treatment strategies for each syndrome. We discuss the various modifications and evolutions of the CRISPR toolkit that expand its therapeutic potential, and other considerations for moving these strategies from bench to bedside. The rapidly growing field of CRISPR therapeutics is providing a myriad of approaches to target a gene, pathway, or transcript for modification. As cures for FXADs have remained elusive, CRISPR opens new avenues to pursue.<\/jats:p>","DOI":"10.3390\/brainsci9010017","type":"journal-article","created":{"date-parts":[[2019,1,22]],"date-time":"2019-01-22T03:08:22Z","timestamp":1548126502000},"page":"17","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["CRISPR to the Rescue: Advances in Gene Editing for the FMR1 Gene"],"prefix":"10.3390","volume":"9","author":[{"given":"Carolyn","family":"Yrigollen","sequence":"first","affiliation":[{"name":"The Raymond G. Perelman Center of Cellular and Molecular Therapeutics, Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beverly","family":"Davidson","sequence":"additional","affiliation":[{"name":"The Raymond G. Perelman Center of Cellular and Molecular Therapeutics, Children\u2019s Hospital of Philadelphia, Philadelphia, PA 19104, USA"},{"name":"Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,1,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/j.bbagen.2009.02.007","article-title":"Fmr1: A gene with three faces","volume":"1790","author":"Oostra","year":"2009","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4948","DOI":"10.1093\/hmg\/ddv216","article-title":"Reversibility of neuropathology and motor deficits in an inducible mouse model for fxtas","volume":"24","author":"Hukema","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1016\/j.neuron.2016.12.016","article-title":"Translation of expanded cgg repeats into fmrpolyg is pathogenic and may contribute to fragile x tremor ataxia syndrome","volume":"93","author":"Sellier","year":"2017","journal-title":"Neuron"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1016\/j.neuron.2013.03.026","article-title":"Cgg repeat-associated translation mediates neurodegeneration in fragile x tremor ataxia syndrome","volume":"78","author":"Todd","year":"2013","journal-title":"Neuron"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1083\/jcb.201010027","article-title":"Spatial code recognition in neuronal rna targeting: Role of rna-hnrnp a2 interactions","volume":"194","author":"Muslimov","year":"2011","journal-title":"J. Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1007\/s00439-017-1807-6","article-title":"Protein sequestration as a normal function of long noncoding rnas and a pathogenic mechanism of rnas containing nucleotide repeat expansions","volume":"136","author":"Morriss","year":"2017","journal-title":"Hum. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1248","DOI":"10.1038\/emboj.2010.21","article-title":"Sam68 sequestration and partial loss of function are associated with splicing alterations in fxtas patients","volume":"29","author":"Sellier","year":"2010","journal-title":"EMBO J."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1111\/nyas.12693","article-title":"Fragile x-associated tremor\/ataxia syndrome","volume":"1338","author":"Hagerman","year":"2015","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"17065","DOI":"10.1038\/nrdp.2017.65","article-title":"Fragile x syndrome","volume":"3","author":"Hagerman","year":"2017","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1016\/j.tig.2018.05.004","article-title":"Gene editing on center stage","volume":"34","author":"Bak","year":"2018","journal-title":"Trends Genet."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using crispr\/cas systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e00471","DOI":"10.7554\/eLife.00471","article-title":"Rna-programmed genome editing in human cells","volume":"2","author":"Jinek","year":"2013","journal-title":"Elife"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1126\/science.1232033","article-title":"Rna-guided human genome engineering via cas9","volume":"339","author":"Mali","year":"2013","journal-title":"Science"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.1038\/s41467-018-04252-2","article-title":"The crispr tool kit for genome editing and beyond","volume":"9","author":"Adli","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.1016\/j.cell.2014.05.010","article-title":"Development and applications of crispr-cas9 for genome engineering","volume":"157","author":"Hsu","year":"2014","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1709","DOI":"10.1126\/science.1138140","article-title":"Crispr provides acquired resistance against viruses in prokaryotes","volume":"315","author":"Barrangou","year":"2007","journal-title":"Science"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1146\/annurev-biophys-062215-010822","article-title":"Crispr-cas9 structures and mechanisms","volume":"46","author":"Jiang","year":"2017","journal-title":"Annu. Rev. Biophys."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2180","DOI":"10.1038\/nprot.2013.132","article-title":"Crispr interference (crispri) for sequence-specific control of gene expression","volume":"8","author":"Larson","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1016\/j.cell.2013.02.022","article-title":"Repurposing crispr as an rna-guided platform for sequence-specific control of gene expression","volume":"152","author":"Qi","year":"2013","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1016\/j.cell.2013.06.044","article-title":"Crispr-mediated modular rna-guided regulation of transcription in eukaryotes","volume":"154","author":"Gilbert","year":"2013","journal-title":"Cell"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1038\/nmeth.2857","article-title":"Efficient genome modification by crispr-cas9 nickase with minimal off-target effects","volume":"11","author":"Shen","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.cell.2014.09.039","article-title":"A protein-tagging system for signal amplification in gene expression and fluorescence imaging","volume":"159","author":"Tanenbaum","year":"2014","journal-title":"Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/nature14136","article-title":"Genome-scale transcriptional activation by an engineered crispr-cas9 complex","volume":"517","author":"Konermann","year":"2015","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1038\/nbt.3199","article-title":"Epigenome editing by a crispr-cas9-based acetyltransferase activates genes from promoters and enhancers","volume":"33","author":"Hilton","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1038\/nbt.3803","article-title":"Increasing the genome-targeting scope and precision of base editing with engineered cas9-cytidine deaminase fusions","volume":"35","author":"Kim","year":"2017","journal-title":"Nat. Biotechnol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/nature17946","article-title":"Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage","volume":"533","author":"Komor","year":"2016","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1216","DOI":"10.1038\/s41591-018-0137-0","article-title":"A high-fidelity cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells","volume":"24","author":"Vakulskas","year":"2018","journal-title":"Nat. Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1126\/science.aad5227","article-title":"Rationally engineered cas9 nucleases with improved specificity","volume":"351","author":"Slaymaker","year":"2016","journal-title":"Science"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"490","DOI":"10.1038\/nature16526","article-title":"High-fidelity crispr-cas9 nucleases with no detectable genome-wide off-target effects","volume":"529","author":"Kleinstiver","year":"2016","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1038\/nature14299","article-title":"In vivo genome editing using staphylococcus aureus cas9","volume":"520","author":"Ran","year":"2015","journal-title":"Nature"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1038\/nature14592","article-title":"Engineered crispr-cas9 nucleases with altered pam specificities","volume":"523","author":"Kleinstiver","year":"2015","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1126\/science.aaq0180","article-title":"Rna editing with crispr-cas13","volume":"358","author":"Cox","year":"2017","journal-title":"Science"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.celrep.2015.08.084","article-title":"Reversion of fmr1 methylation and silencing by editing the triplet repeats in fragile x ipsc-derived neurons","volume":"13","author":"Park","year":"2015","journal-title":"Cell Rep."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Xie, N., Gong, H., Suhl, J.A., Chopra, P., Wang, T., and Warren, S.T. (2016). Reactivation of fmr1 by crispr\/cas9-mediated deletion of the expanded cgg-repeat of the fragile x chromosome. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0165499"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1016\/j.cell.2018.01.012","article-title":"Rescue of fragile x syndrome neurons by DNA methylation editing of the fmr1 gene","volume":"172","author":"Liu","year":"2018","journal-title":"Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"282","DOI":"10.3389\/fnmol.2018.00282","article-title":"Targeted reactivation of fmr1 transcription in fragile x syndrome embryonic stem cells","volume":"11","author":"Haenfler","year":"2018","journal-title":"Front. Mol. Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1038\/jhg.2009.121","article-title":"Examination of fmr1 transcript and protein levels among 74 premutation carriers","volume":"55","author":"Peprah","year":"2010","journal-title":"J. Hum. Genet."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1111\/j.1399-0004.2011.01675.x","article-title":"Fragile x-associated tremor\/ataxia phenotype in a male carrier of unmethylated full mutation in the fmr1 gene","volume":"82","author":"Loesch","year":"2012","journal-title":"Clin. Genet."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1111\/cge.12278","article-title":"Fxtas in an unmethylated mosaic male with fragile x syndrome from chile","volume":"86","author":"Pugin","year":"2014","journal-title":"Clin. Genet."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1038\/s41551-018-0252-8","article-title":"Nanoparticle delivery of crispr into the brain rescues a mouse model of fragile x syndrome from exaggerated repetitive behaviours","volume":"2","author":"Lee","year":"2018","journal-title":"Nat. Biomed. Eng."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"748","DOI":"10.1016\/j.msec.2017.05.110","article-title":"Effect of acute and long-term administration of gold nanoparticles on biochemical parameters in rat brain","volume":"79","author":"Ferreira","year":"2017","journal-title":"Mater. Sci. Eng. C Mater. Biol. Appl."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2227","DOI":"10.1016\/j.celrep.2018.02.014","article-title":"A single administration of crispr\/cas9 lipid nanoparticles achieves robust and persistent in vivo genome editing","volume":"22","author":"Finn","year":"2018","journal-title":"Cell Rep."},{"key":"ref_43","unstructured":"Lonser, R., Sarntinoranont, M., and Bankiewicz, K. (2019). Crispr-cas gene editing for neurological disease. Nervous System Drug Delivery: Principles and Practice, Academic Press\/Elsevier. in press."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.ymthe.2016.11.010","article-title":"Crispr\/cas9 editing of the mutant huntingtin allele in vitro and in vivo","volume":"25","author":"Monteys","year":"2017","journal-title":"Mol. Ther"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/gt.2016.89","article-title":"Deletion of the gaa repeats from the human frataxin gene using the crispr-cas9 system in yg8r-derived cells and mouse models of friedreich ataxia","volume":"24","author":"Ouellet","year":"2017","journal-title":"Gene Ther."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1038\/nbt.2908","article-title":"Dimeric crispr rna-guided foki nucleases for highly specific genome editing","volume":"32","author":"Tsai","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1080\/07388551.2017.1378999","article-title":"Inducible crispr genome-editing tool: Classifications and future trends","volume":"38","author":"Dai","year":"2018","journal-title":"Crit. Rev. Biotechnol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.pneurobio.2018.04.008","article-title":"In vivo methods for acute modulation of gene expression in the central nervous system","volume":"168","author":"Cwetsch","year":"2018","journal-title":"Prog. Neurobiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1007\/s40259-017-0234-5","article-title":"Adeno-associated virus (aav) as a vector for gene therapy","volume":"31","author":"Naso","year":"2017","journal-title":"BioDrugs"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1126\/science.aad5143","article-title":"In vivo genome editing improves muscle function in a mouse model of duchenne muscular dystrophy","volume":"351","author":"Nelson","year":"2016","journal-title":"Science"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1038\/nmeth.4278","article-title":"Circle-seq: A highly sensitive in vitro screen for genome-wide crispr-cas9 nuclease off-targets","volume":"14","author":"Tsai","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1186\/s13059-015-0817-8","article-title":"Characterization of staphylococcus aureus cas9: A smaller cas9 for all-in-one adeno-associated virus delivery and paired nickase applications","volume":"16","author":"Friedland","year":"2015","journal-title":"Genome Biol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3100","DOI":"10.1038\/npp.2014.167","article-title":"Reduced phenotypic severity following adeno-associated virus-mediated fmr1 gene delivery in fragile x mice","volume":"39","author":"Gholizadeh","year":"2014","journal-title":"Neuropsychopharmacology"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Aschauer, D.F., Kreuz, S., and Rumpel, S. (2013). Analysis of transduction efficiency, tropism and axonal transport of aav serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0076310"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1038\/mt.2009.286","article-title":"Comparative transduction efficiency of aav vector serotypes 1-6 in the substantia nigra and striatum of the primate brain","volume":"18","author":"Markakis","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1038\/gt.2012.20","article-title":"Directed evolution of novel adeno-associated viruses for therapeutic gene delivery","volume":"19","author":"Bartel","year":"2012","journal-title":"Gene Ther."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/s41434-018-0005-z","article-title":"Widespread transduction of astrocytes and neurons in the mouse central nervous system after systemic delivery of a self-complementary aav-php.B vector","volume":"25","author":"Rincon","year":"2018","journal-title":"Gene Ther."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1038\/cr.2017.16","article-title":"A non-viral crispr\/cas9 delivery system for therapeutically targeting hbv DNA and pcsk9 in vivo","volume":"27","author":"Jiang","year":"2017","journal-title":"Cell Res."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1002\/anie.201610209","article-title":"Non-viral crispr\/cas gene editing in vitro and in vivo enabled by synthetic nanoparticle co-delivery of cas9 mrna and sgrna","volume":"56","author":"Miller","year":"2017","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1012","DOI":"10.1101\/gr.171322.113","article-title":"Highly efficient rna-guided genome editing in human cells via delivery of purified cas9 ribonucleoproteins","volume":"24","author":"Kim","year":"2014","journal-title":"Genome Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.jns.2015.03.031","article-title":"Fragile x premutation carriers: A systematic review of neuroimaging findings","volume":"352","author":"Brown","year":"2015","journal-title":"J. Neurol. Sci."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"4904","DOI":"10.1002\/hbm.22521","article-title":"Topological methods reveal high and low functioning neuro-phenotypes within fragile x syndrome","volume":"35","author":"Romano","year":"2014","journal-title":"Hum. Brain Mapp."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/2040-2392-2-2","article-title":"Neuropathologic features in the hippocampus and cerebellum of three older men with fragile x syndrome","volume":"2","author":"Greco","year":"2011","journal-title":"Mol. Autism"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"5561","DOI":"10.2147\/IJN.S149022","article-title":"Theranostic applications of nanoparticles in neurodegenerative disorders","volume":"13","author":"Ramanathan","year":"2018","journal-title":"Int. J. Nanomed."},{"key":"ref_65","first-page":"280813","article-title":"Early intervention combined with targeted treatment promotes cognitive and behavioral improvements in young children with fragile x syndrome","volume":"2012","author":"Winarni","year":"2012","journal-title":"Case Rep. Genet."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1038\/nbt.3117","article-title":"Guide-seq enables genome-wide profiling of off-target cleavage by crispr-cas nucleases","volume":"33","author":"Tsai","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1038\/nbt.3101","article-title":"Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases","volume":"33","author":"Frock","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/nbt.3127","article-title":"Unbiased detection of off-target cleavage by crispr-cas9 and talens using integrase-defective lentiviral vectors","volume":"33","author":"Wang","year":"2015","journal-title":"Nat. Biotechnol"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1038\/nmeth.3284","article-title":"Digenome-seq: Genome-wide profiling of crispr-cas9 off-target effects in human cells","volume":"12","author":"Kim","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1101\/gr.199588.115","article-title":"Genome-wide target specificities of crispr-cas9 nucleases revealed by multiplex digenome-seq","volume":"26","author":"Kim","year":"2016","journal-title":"Genome Res."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"600","DOI":"10.1038\/nmeth.4284","article-title":"Mapping the genomic landscape of crispr-cas9 cleavage","volume":"14","author":"Cameron","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1038\/nmeth.2408","article-title":"Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing","volume":"10","author":"Crosetto","year":"2013","journal-title":"Nat. Methods"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"15058","DOI":"10.1038\/ncomms15058","article-title":"Bliss is a versatile and quantitative method for genome-wide profiling of DNA double-strand breaks","volume":"8","author":"Yan","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.biotechadv.2016.12.003","article-title":"Detection of on-target and off-target mutations generated by crispr\/cas9 and other sequence-specific nucleases","volume":"35","author":"Zischewski","year":"2017","journal-title":"Biotechnol. Adv."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"e118","DOI":"10.1093\/nar\/gkv575","article-title":"Cas9-chromatin binding information enables more accurate crispr off-target prediction","volume":"43","author":"Singh","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1038\/nmeth.3543","article-title":"Crisprscan: Designing highly efficient sgrnas for crispr-cas9 targeting in vivo","volume":"12","author":"Vejnar","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1007\/s12539-018-0298-z","article-title":"Review of crispr\/cas9 sgrna design tools","volume":"10","author":"Cui","year":"2018","journal-title":"Interdiscip Sci."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1186\/s13059-016-1012-2","article-title":"Evaluation of off-target and on-target scoring algorithms and integration into the guide rna selection tool crispor","volume":"17","author":"Haeussler","year":"2016","journal-title":"Genome Biol."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1038\/nbt.3290","article-title":"Chemically modified guide rnas enhance crispr-cas genome editing in human primary cells","volume":"33","author":"Hendel","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"2980","DOI":"10.1016\/j.celrep.2017.08.075","article-title":"The self-inactivating kamicas9 system for the editing of cns disease genes","volume":"20","author":"Merienne","year":"2017","journal-title":"Cell Rep."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1038\/s41467-017-00124-3","article-title":"Inactivation of pol theta and c-nhej eliminates off-target integration of exogenous DNA","volume":"8","author":"Zelensky","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.celrep.2014.02.040","article-title":"Quantifying genome-editing outcomes at endogenous loci with smrt sequencing","volume":"7","author":"Hendel","year":"2014","journal-title":"Cell Rep."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"3964","DOI":"10.1038\/ncomms4964","article-title":"Engineering human tumour-associated chromosomal translocations with the rna-guided crispr-cas9 system","volume":"5","author":"Torres","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.tibtech.2014.12.001","article-title":"Quantifying on- and off-target genome editing","volume":"33","author":"Hendel","year":"2015","journal-title":"Trends Biotechnol."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"318","DOI":"10.3389\/fgene.2014.00318","article-title":"Clinical and molecular implications of mosaicism in fmr1 full mutations","volume":"5","author":"Pretto","year":"2014","journal-title":"Front. Genet."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1136\/jmedgenet-2013-102021","article-title":"Cgg allele size somatic mosaicism and methylation in fmr1 premutation alleles","volume":"51","author":"Pretto","year":"2014","journal-title":"J. Med. Genet."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/nbt.4207","article-title":"Unexpected crispr on-target effects","volume":"36","author":"Lee","year":"2018","journal-title":"Nat. Biotechnol."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.jmb.2018.05.044","article-title":"Crispr ethics: Moral considerations for applications of a powerful tool","volume":"431","author":"Brokowski","year":"2019","journal-title":"J. Mol. Biol."}],"container-title":["Brain Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3425\/9\/1\/17\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:27:39Z","timestamp":1760185659000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3425\/9\/1\/17"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,1,21]]},"references-count":88,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,1]]}},"alternative-id":["brainsci9010017"],"URL":"https:\/\/doi.org\/10.3390\/brainsci9010017","relation":{},"ISSN":["2076-3425"],"issn-type":[{"value":"2076-3425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,1,21]]}}}