02081nam a2200253Ia 4500 MX-MdCICY 20260521091537.0 CICY B-16781 CRISPR/Cas9-Enabled Multiplex Genome Editing and Its Application Progress in Molecular Biology and Translational Science, 149, p.111-132, 2017 The CRISPR/Cas9 system is a prevalent and versatile genome-editing tool of choice for basic and applied biological research. An exchange of a 20-bp spacer sequence in the gRNA can easily reprogram Cas9 to target a different DNA site. By expressing or providing multiple gRNAs, the system also enables multiplex genome editing at high efficiencies. Current approaches for providing multiple gRNAs in vivo include the use of multigene cassettes to express several gRNAs, Csy4-based excision, arrays of crRNAs, ribozyme-flanked gRNAs, tRNA-dependent cleavage of gRNAs, and direct introduction of Cas9 proteins preloaded with different gRNAs. By simultaneously targeting multiple DNA sequences, multiplex genome editing can be used to knockout multiple genes or delete chromosomal fragments. Off-target risk can also be reduced by Cas9-dimers that require the simultaneous expression of two gRNAs. With multiple gRNAs, specific gene expression or methylation status can be efficiently controlled by dCas9 fused to activators, repressors, methyltransferase, demethylase, or other functional domains. As a result, multiplex genome editing is expected to accelerate functional discovery of plant genes as well as genetic improvement of agricultural crops. CRISPR/CAS GENE EDITING GENOME EDITING GUIDE RNA EXPRESSION MULTIPLEX EDITING Minkenberg, B. Wheatley, M. Yang, Y. https://drive.google.com/file/d/1sMb3SjMZKU9M2vkRNBdO9Cd_rEA0guM7/view?usp=drivesdk Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx Loc REF1 250602s9999 xx |||||s2 |||| ||und|d 26887 26887 0 0 Loc 0 0 F1 CICY CICY RE 2025-06-25 0 B-16781 2025-06-25 16:01:58 2025-06-25 REF1