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Heritable and precise zebrafish genome editing using a CRISPR-Cas system


, : Heritable and precise zebrafish genome editing using a CRISPR-Cas system. Plos One 8(7): E68708-E68708

We have previously reported a simple and customizable CRISPR (clustered regularly interspaced short palindromic repeats) RNA-guided Cas9 nuclease (RGN) system that can be used to efficiently and robustly introduce somatic indel mutations in endogenous zebrafish genes. Here we demonstrate that RGN-induced mutations are heritable, with efficiencies of germline transmission reaching as high as 100%. In addition, we extend the power of the RGN system by showing that these nucleases can be used with single-stranded oligodeoxynucleotides (ssODNs) to create precise intended sequence modifications, including single nucleotide substitutions. Finally, we describe and validate simple strategies that improve the targeting range of RGNs from 1 in every 128 basepairs (bps) of random DNA sequence to 1 in every 8 bps. Together, these advances expand the utility of the CRISPR-Cas system in the zebrafish beyond somatic indel formation to heritable and precise genome modifications.

US$19.90

PMID: 23874735

DOI: 10.1371/journal.pone.0068708


Other references

Zemanek, J., 1970: Effect of environmental factors and some chemicals on germination and emergence of the weed Tripleurospermum maritimum (L.) Sch. Bip. [245.998:132.2:224].In laboratory experiments seeds germinated best at 15-28 deg C, with a moisture content of < 20% and 15-30% in soil and sand respectively and when surface sown. Germination was stimulated by NH4-N and particularly by No3-N,...

Sprecher D.L.; Pierce J.S.; Kruth H.; Alfieris G.; Lakatos E.; Brewer H.B.; Epstein S.E.; Patterson R.E., 1985: Influence of a hypercholesterolemic diet on the coronary vasodilator response to histamine in miniature pigs. American Heart Association Monograph (114): III-76

Boltrukevich, S.I.; Kalygin, A.V., 1989: Osteoplasty in wound infection. The outcome of a surgical intervention in the purulent wound together with other common clinical factors depends to a determining degree on the quality of the plastic material and its resistance to infection. The authors used allogenic transplants...

Yusakul, G.; Putalun, W.; Udomsin, O.; Juengwatanatrakul, T.; Chaichantipyuth, C., 2011: Comparative analysis of the chemical constituents of two varieties of Pueraria candollei. A high-performance liquid chromatography (HPLC) method was developed to determine the contents of miroestrol and deoxymiroestrol in the tubers of Pueraria candollei var. mirifica and P. candollei var. candollei. The linear detection ranges were 0....

Sullivan, C.H.; Norman, J.T.; Borrás, T.; Grainger, R.M., 1989: Developmental regulation of hypomethylation of delta-crystallin genes in chicken embryo lens cells. Sequences in the two delta-crystallin genes become hypomethylated when they are expressed in the chick lens. This system is particularly advantageous for studying temporal changes in hypomethylation, since lens tissue can be isolated at all develo...

Reddy, C.R.; Narasaiah, I.L.; Parvathi, G., 1969: Epidemiological studies on guinea-worm infection. The 10, 000 inhabitants of 4 villages in South India were interviewed and the prevalence of Dracunculus medinensis infection was found to vary from 11 % to 54% and to increase with the proportion of the population using step-wells. Questioning rev...

Barbakow, F.H.; Cleaton-Jones, P.; Friedman, D., 1980: An evaluation of 566 cases of root canal therapy in general dental practice. 2. Postoperative observations. Journal of Endodontics 6(3): 485-489

Park, C.; Raigrodski, A.J.; Rosen, J.; Spiekerman, C.; London, R.M., 2009: Accuracy of implant placement using precision surgical guides with varying occlusogingival heights: an in vitro study. Surgical guides may interfere with effective use of surgical instrumentation during implant placement in the posterior segments where interocclusal distance may be limited. The purpose of this study was to measure and compare the accuracy of poste...

Fernandes, A.J.; Diotaiuti, L.; Dias, J.C.; Romanha, A.J.; Chiari, E., 1994: Relationships between Trypanosoma cruzi transmission cycles in the county of Bambuí, Minas Gerais, Brazil. This study examines recent relationships between domestic and sylvatic transmission cycles of T. cruzi in the county of Bambuí, MG, Brazil. In the late l930s, Panstrongylus megistus was found in 75% of houses. Subsequently, Triatoma infestans bec...

Howes Jones D.; Barlow J.C., 1988: The structure of the call note system of the warbling vireo. This study presents a detailed description of the structure of the call note system of the Warbling Vireo, Vireo gilvus gilvus, in southern Ontario. Properties or attributes used to describe calls are form, context, message, frequency of occurrenc...