A CRISPer Guide for Generating Superior sgRNA Libraries

The ability to design and synthesize any DNA sequence is at the core of all synthetic biology efforts. Agilent’s capability to synthesize up to 2.4 x 10⁵ high quality DNA oligomers of 200 nt in length on a single chip enables a variety of experimental approaches otherwise not accessible. Examples of applications include sequence selection prior to high throughput sequencing applications, low cost synthesis of large DNA sequences, and targeted high throughput mutagenesis. Another application is the generation of complex plasmid libraries containing short, defined sequence variants. One example is the construction of libraries of short guide RNAs for CAS9 to enable genome-wide gene knock out screening or CAS mediated transcriptional activation or repression (CRISPR-A/CRISPR-I). We have developed novel chemistries that utilize an overlap-based cloning method and workflows necessary for generating highest quality, low-bias, low-skew plasmid libraries essential for successful use of these libraries.