Titlebook: CRISPR Gene Editing; Methods and Protocol Yonglun Luo Book 2019 Springer Science+Business Media, LLC, part of Springer Nature 2019 CRISPR-C

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Rapid and Efficient Gene Deletion by CRISPR/Cas9Here we describe our method for rapid and efficient generation of gene knockout or deletion cells using CRISPR/Cas9 within the time span of one month. The design of gRNAs, plasmid cloning, transfection, cell culturing, positive clone selection, and screening can be obtained from this method.

fastness

Rapid Quantitative Evaluation of CRISPR Genome Editing by TIDE and TIDERorithm (available at . or .). The routine is easy, fast, and provides much more detailed information than current enzyme-based assays. TIDE and TIDER accelerate testing and designing of DSB-based genome editing strategies.

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Fast and Quantitative Identification of Ex Vivo Precise Genome Targeting-Induced Indel Events by IDAf CRISPR/Cas9 KO and repurposed systems. To this end, validation of gRNA functionality depends on efficient, accurate, and sensitive identification of indels induced by a given gRNA design. For in vitro indel profiling the most commonly used methods are based on amplicon size discrimination or seque

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Functional Evaluation of CRISPR Activity by the Dual-Fluorescent Surrogate System: C-Checktivity, as well as for enrichment of gene edited cells. In this chapter, we will give a step-by-step instruction on the design, generation, and application of the C-Check system for quantifying gRNA activities.

Deceit

Rapid and Simple Screening of CRISPR Guide RNAs (gRNAs) in Cultured Cells Using Adeno-Associated Virspecificities. Here, we describe a fast and simple protocol for small-scale packaging of CRISPR/Cas9 components into AAV vectors. To showcase its potential, we employ this method for screening of gRNAs targeting the murine miR-122 locus in Hepa1–6 cells (using AAV serotype 6, AAV6) or the 5′LTR of t

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interior

https://doi.org/10.1007/978-3-030-89525-9specificities. Here, we describe a fast and simple protocol for small-scale packaging of CRISPR/Cas9 components into AAV vectors. To showcase its potential, we employ this method for screening of gRNAs targeting the murine miR-122 locus in Hepa1–6 cells (using AAV serotype 6, AAV6) or the 5′LTR of t