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Jia, Y., Xu, R.G., Ren, X., Ewen-Campen, B., Rajakumar, R., Zirin, J., Yang-Zhou, D., Zhu, R., Wang, F., Mao, D., Peng, P., Qiao, H.H., Wang, X., Liu, L.P., Xu, B., Ji, J.Y., Liu, Q., Sun, J., Perrimon, N., Ni, J.Q. (2018). Next-generation CRISPR/Cas9 transcriptional activation in Drosophila using flySAM.  Proc. Natl. Acad. Sci. U.S.A. 115(18): 4719--4724.
FlyBase ID
FBrf0238848
Publication Type
Research paper
Abstract

CRISPR/Cas9-based transcriptional activation (CRISPRa) has recently emerged as a powerful and scalable technique for systematic overexpression genetic analysis in Drosophila melanogaster We present flySAM, a potent tool for in vivo CRISPRa, which offers major improvements over existing strategies in terms of effectiveness, scalability, and ease of use. flySAM outperforms existing in vivo CRISPRa strategies and approximates phenotypes obtained using traditional Gal4-UAS overexpression. Moreover, because flySAM typically requires only a single sgRNA, it dramatically improves scalability. We use flySAM to demonstrate multiplexed CRISPRa, which has not been previously shown in vivo. In addition, we have simplified the experimental use of flySAM by creating a single vector encoding both the UAS:Cas9-activator and the sgRNA, allowing for inducible CRISPRa in a single genetic cross. flySAM will replace previous CRISPRa strategies as the basis of our growing genome-wide transgenic overexpression resource, TRiP-OE.

PubMed ID
PubMed Central ID
PMC5939105 (PMC) (EuropePMC)
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Secondary IDs
    Language of Publication
    English
    Additional Languages of Abstract
    Parent Publication
    Publication Type
    Journal
    Abbreviation
    Proc. Natl. Acad. Sci. U.S.A.
    Title
    Proceedings of the National Academy of Sciences of the United States of America
    Publication Year
    1915-
    ISBN/ISSN
    0027-8424
    Data From Reference
    Alleles (71)
    Genes (31)
    Molecular Constructs (3)
    Natural transposons (1)
    Insertions (5)
    Experimental Tools (7)
    Transgenic Constructs (54)