CRISPR System with AAV

SpCas9
Gene-Knockout
Zhang’s group from the Broad Institute (MIT) breed a SpCas9 knock-in mouse that can express SpCas9 constitutively or Cre-dependently in cells of whole body, and demonstrated that utilizing AAV delivering sgRNA and donor sequence can precisely modify the KRAS gene into a tumor-causing mutation, which then results in rapid tumor growth in 1 to 3 months (5). SpCas9 mouse is now available from Jackson Lab.



Gene-Activation


SaCas9

On the final day of 2015, three groups published their promising results on utilizing AAV to deliver Cas9 to therapy in DMD mouse as a therapeutics published in Science (8, 9, and 10). All three groups applied the same strategy: injection of AAV expressing SaCas9/SpCas9 and sgRNAs to delete the mutated Exon 23 in DMD mouse model, thereby restoring the truncated but still functional Dystrophin.

Refferences
2. Le Cong et al. Science 339,819 (2013); DOI: 10.1126/science.1231143
3. Prashant Mali et al. Science 339,823 (2013); DOI: 10.1126/science.1232033
4. Randall J. Platt et al. Cell 159, 1–16 (2014); DOI: 10.1016/j.cell.2014.09.014
5. F. Ann Ran et al. Nature 520, 186 (2015); doi:10.1038/nature14299
6. Florian Schmidt and Dirk Grimm. Biotechnology J. 10, 258 (2015); DOI0.1002/biot.201400529
7. David B. T. Cox et al. Nature Medicine 21,121 (2015); doi: 10.1038/nm.3793.
8. C. Long et al., Science 10.1126/science.aad5725 (2015)
9. C. E. Nelson et al., Science 10.1126/science.aad5143 (2015)
10. M. Tabebordbar et al., Science 10.1126/science.aad5177 (2015).
11. Dahlman JE et al., Nat Biotechnol. 2015 Nov;33(11):1159-61.
12. Nishimasu et al., Cell 162, 1113–1126 (2015)