Titlebook: CRISPR-Cas Methods; Volume 2 M. Tofazzal Islam,Kutubuddin Ali Molla Book 2021 The Editor(s) (if applicable) and The Author(s), under exclus

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https://doi.org/10.1007/978-3-662-08919-4al applications. Here, we describe the design principle of the microRNA-responsive Cas9 and AcrllA4 (anti-CRISPR protein for SpCas9) switch and the procedures for executing its cell-type-specific genome editing.

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Generating Clonal Seeds from Hybrid Rice with CRISPR-Cas9, the . gene induces formation of maternal haploid seeds. Genome editing of all these four genes in hybrid rice simultaneously could fix the heterozygosity and obtain clonal seeds from hybrid rice. Here, we describe a detailed method for generating clonal seeds from hybrid rice by using the multiplex CRISPR-Cas9 technology.

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An Approach to Proximity Ligation by T4 RNA Ligase to Screen sRNA That Regulate CRISPR-Cas Systems,RNA ligase 1 (single-stranded RNA ligase) to link two base-paired RNA molecules to investigate potential interaction between sRNA and CRISPR-Cas system. Our goal is to provide readers a detailed method of identifying candidate sRNAs that may regulate CRISPR-Cas adaptation and/or other functions through unbiased screening and validation.

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Naturheilkundliche Pflege von Kindern structure in target recognition and efficacy of CRISPR-Cas systems, it is vital to assess the gRNA secondary structure. Here, we describe a protocol to determine the gRNA secondary structure using RNA-fold software and explain how to interpret the results.

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Zwischenbilanz und Perspektiven,. Here, we describe a simple method to accurately assemble completely natural, multiplex CRISPR arrays that can be completed in 1–2 days. This should be of great use both in prokaryotes with their own native CRISPR systems and in eukaryotes when paired with Cas12a or other CRISPR nucleases that also process their own arrays.

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Chr. Gutenbrunner,G. Hildebrandtposed of nCas9, a rat cytidine deaminase enzyme APOBEC1, and a uracil DNA glycosylase inhibitor (UGI). The CBE was constructed in the PTF101 vector background. The PTF101-CBE has successfully achieved single-base alteration in soybean. Here, we present a detailed protocol of this base editor to generate a point mutation in soybean.

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