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There are practical considerations to bear in mind with regard to reagents and materials for the design and execution of in vivo CRISPR screens. Due to the limited number of cells that can be injected into each animal, it is challenging to perform genome‐wide In vivo screens with appropriate library coverage. Therefore, most researchers have chosen to use a focused mini‐pool gRNA library to ensure high screen quality. While off‐the‐shelf sub‐pool gRNA libraries are available from major commercial vendors, custom‐designed gRNA libraries can be constructed by CROs like Merck, Horizon Discovery, and GenScript. To make your own gRNA libraries, Twist Bioscience, CustomArray (now part of GenScript), Agilent, and IDT are the major suppliers for oligonucleotide library synthesis. It is easier to conduct pooled in vivo CRISPR screens using cells from transgenic mice constitutively expressing Cas9, as this bypasses the need for endonuclease delivery/selection into target cells. Several different Cas9 knock‐in mice are available from The Jackson Laboratory, but licenses for research use must be obtained from the original inventors for industry/biotech institutions. T cell receptor transgenic mice from The Jackson Laboratory and other suppliers are commonly used for adoptive T cell transfer experiments, which allow for T‐cell‐based in vivo CRISPR screening. Major CROs that offer in vivo genetic screens are included in Table 4.2.

Table 4.2 Genome editing service providers.