Titlebook: In Vitro Mutagenesis; Methods and Protocol Andrew Reeves Book 2017 Springer Science+Business Media New York 2017 gene and genome editing.fa

壓迫

Apogee

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Design and Validation of CRISPR/Cas9 Systems for Targeted Gene Modification in Induced Pluripotent Sne the procedures for design, screening, and validation of CRISPR/Cas9 systems for targeted modification of coding sequences in the human genome and how to perform genome editing in induced pluripotent stem cells with high efficiency and specificity.

懲罰

Use of CRISPR/Cas Genome Editing Technology for Targeted Mutagenesis in Riceic important traits have been rapidly generated, which indicates the potential applications in both scientific research and plant breeding. In this chapter, we describe a standard gene-editing procedure to effectively target rice genes and to make specific rice mutants using the CRISPR/Cas9 system mediated by . transformation.

殺菌劑

Generation of Stable Knockout Mammalian Cells by TALEN-Mediated Locus-Specific Gene Editingnd generation of TALENs using serial and hierarchical digestion and ligation steps, and using the synthesized TALEN pairs to achieve locus-specific targeted gene editing in mammalian cell lines using a modified clonal selection strategy in an easy and cost-efficient manner.

organism

Use of Group II Intron Technology for Targeted Mutagenesis in ,utants in the obligate, intracellular bacterial pathogen .. The methods employed for intron targeting, mutant selection, and mutant verification will be outlined including available selection markers, gene targeting strategies, and potential pitfalls.

戰(zhàn)勝

Efficient Generation of Gene-Modified Mice by Haploid Embryonic Stem Cell-Mediated Semi-cloned TechnCs injection (ICAHCI), which provides a new route to obtain genetically modified mice. In this chapter, we describe the procedures for AG-haESCs culturing, enrichment of haploid cells by FACS, genomic manipulation in DKO-AG-haESCs by CRISPR/Cas9 and generation of live SC mice with gene-modified DKO-AG-haESCs.

意外的成功

Mutagenesis and Genome Engineering of Epstein–Barr Virus in Cultured Human Cells by CRISPR/Cas9artificial chromosome-based recombineering. This approach might also prove useful in the cure of EBV infection. In this chapter, we use the knockout of the BART promoter as an example to detail the experimental procedures for construction of recombinant EBV in human cells.

善變

Development of CRISPR/Cas9 for Efficient Genome Editing in ,ntation. The technical details of the strategy, including CRISPR plasmid construction, target construct generation, parasite transfection and positive clone identification are also provided. These methods are easy to customize to any gene of interest (GOI) and will greatly accelerate studies on this important pathogen.

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