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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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Generation of Knockout and Fragment Deletion Mutants in Soybean by CRISPR-Cas9,ng. CRISPR-Cas systems can generate highly specific double-strand breaks (DSBs) at the target site, and desired sequence modifications can be introduced during the DSB repair process, such as nonhomologous end-joining (NHEJ) or homology-directed repair (HDR) pathways. Among Cas nuclease proteins, Ca
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Targeted Base Editing in Soybean Using a CRISPR-Cas9 Cytidine Deaminase Fusion,ingle-base change or SNP without double-strand break in the genome. Cytosine base editors (CBEs) and adenine base editors (ABEs) are two types of base editors, which are composed of a deactivated Cas9 endonuclease (dCas9) or a Cas9 nickase (nCas9) and a catalytic domain which is capable of deaminati
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Efficient CRISPR-Cas9-Mediated Genome Editing in Tomato,-Cas system has become commonplace around the world. This system has been adopted in several plant species. In this protocol, we describe, in a step-wise manner, guidelines for the selection of a suitable vector and target sites, design of sgRNAs, including analysis of off-target activity, construct
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Cell-Type-Specific CRISPR-Cas9 System with miRNAs,e. Regulating Cas9 activity is necessary for the system’s precise genome editing, with editing in only specific cell types being key for future clinical applications. Here, we describe the design principle of the microRNA-responsive Cas9 and AcrllA4 (anti-CRISPR protein for SpCas9) switch and the pr
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CRISPR/Cas9 Gene Editing in Mammalian Cells Using LentiCRISPRv2/LentiGuide-Puro Vectors,l as biotechnology. CRISPR/Cas9 is a form of bacterial defense mechanism that can be used for editing genomes by targeting a 20-nucleotide sequence using a guide RNA and nuclease enzyme called Cas9 enzyme that cleaves target gene. Different protocols have been provided; however, the success of CRISP
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