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Current Strategies for Increasing Knock-In Efficiency in CRISPR Cas9 . . . Strategies for increasing the knock-in efficiency in CRISPR Cas9-based genome editing Increasing CRISPR Cas9-mediated genome editing events involves factors such as the type of cell (a), as well as indirect (b) or direct (c) HDR modulation by using several strategies
Principles of CRISPR-Cas9 technology: Advancements in genome editing . . . The rapid advancement of CRISPR-Cas9 technology has instigated a profound transformation in genome editing with significant implications for fields like health, agriculture, and biotechnology This review provides an overview of the historical significance and fundamental components of CRISPR-Cas9, notably the Cas9 protein and guide RNA, underscoring its pivotal role in genetic manipulation
Telomerase reverse transcriptase gene knock‐in unleashes enhanced . . . The Tert knock-in (TertKI) mice harboring the EF1α-Tert gene displayed elevated telomerase activity, elongated telomeres, and extended lifespan, with no spontaneous genotoxicity or carcinogenicity The TertKI mice showed also enhanced wound healing, characterized by significantly increased expression of Fgf7, Vegf, and collagen
CRISPR Cas9-Mediated Gene Knockout and Knockin Human iPSCs Here, we provide a protocol to successfully generate gene knockout and or knockin iPSCs We include detailed information on the design of guide RNAs (gRNAs), T7 endonuclease assay to detect on-target CRISPR Cas9 editing events, DNA electroporation of the iPSCs with a ribonucleoprotein complex, and single-cell cloning steps for the selection of
Current Strategies for Increasing Knock-In Efficiency in CRISPR Cas9 . . . Since its discovery in 2012, the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) system has supposed a promising panorama for developing novel and highly precise genome editing-based gene therapy (GT) alternatives, leading to overcoming the challenges associated with classical GT Classical GT aims to deliver transgenes to the cells via
The application of CRISPR Cas9–based genome-wide screening to disease . . . This technology has become a cornerstone for genome-wide screenings in disease research This review offers a comprehensive examination of how CRISPR Cas9-based genetic screening has been leveraged to uncover genes that play a role in disease mechanisms, focusing on areas such as cancer development and viral replication processes