GENOME EDITING 113
approach should you choose? To generate a single- or double- knockout quickly, try CRISPR/ Cas9.
Otherwise, TALEN offers fewer off-target effects and target sequence requirements. ES cell gene targeting remains important for the generation of conditional and inducible alleles.
Cyagen Biosciences is a contract research organisation that generates custom, genetically modified mouse and rat models by ES cell homologous recombination, TALEN, and CRISPR/Cas9 methods.
Te organisation serves pharmaceutical companies and academic bioscience research labs worldwide. Its range of services include mouse and rat transgenic, knockout, knockin, humanisation, and nuclease-mediated genome editing.
Fig. 3. Cyagen employs three full-time microinjectionists who perform blastocyst and pronuclear injections for new animal models.
For more information ✔ at
www.scientistlive.com/eurolab
REFERENCES: 1
based methods for genome engineering. Trends Biotechnol. 31: 397-405 2
Cas9 knockout screening in human cells. Science 343: 84-87 3
nucleases in human cells. Nat Biotechnol. 31: 822-826. 4
Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD (2013) High-frequency off-target mutagenesis induced by CRISPR-Cas
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 5
Fig. 4. A chimera from traditional gene targeting. TALEN and CRISPR/Cas9 technology bypasses this stage, saving months.
nickases. Genome Res. 24: 132-141 6
279-284 7
Cho SW, Kim S, Kim Y, Kweon J, Kim HS, Bae S, Kim JS (2014) Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and
Fu Y, Sander JD, Reyon D, Cascio VM, Joung JK (2014) Improving CRISPR- Cas nuclease specificity using truncated guide RNA. Nat Biotechnol. 32:
Park CY, Kim J, Kweon J, Son JS, Lee JS, Yoo JE, Cho SR, Kim JH, Kim JS,
Kim DW (2014) Targeted inversion and reversion of the blood coagulation factor 8 gene in human iPS cells using TALENs. Proc Natl Acad Sci USA 111:
9253-9258 8
Tesson L, Usal C, Ménoret S, Leung E, Niles BJ, Remy S, Santiago Y,
Vincent AI, Meng X, Zhang L, Gregory PD, Anegon I, Cost GJ (2011) Knockout rats generated by embryo microinjection of TALENs. Nat
Biotechnol. 29: 695-696 9
Hwang WY, Fu Y, Reyon D, Maeder ML, Tsai SQ, Sander JD, Peterson
RT, Yeh JR, Joung JK (2013) Efficient genome editing in zebrafish using a CRISPR-Cas system. Nat Biotechnol. 31: 227-229
Fig. 5. Cyagen TALEN or CRISPR/Cas9 knockout and knockin mice are available on the C57BL/6 (shown) or FVB background.
Marvin Yingbin Ouyang is with Cyagen Biosciences in the USA.
www.cyagen.com
www.scientistlive.com
Gaj T, Gersbach CA, Barbas CF 3rd (2013) ZFN, TALEN, and CRISPR/Cas- 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-
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