A Toolkit for Rapid Modular Construction of Biological Circuits in Mammalian Cells

João Pedro Fonseca; Alain R. Bonny; G. Renuka Kumar; Andrew H. Ng; Jason Town; Qiu Chang Wu; Elham Aslankoohi; Susan Y. Chen; Patrick Harrigan; Lindsey C. Osimiri; Amy L. Kistler; Hana El-Samad

Molecular cloning is the cornerstone of modern biological research. It is required to generate DNA vectors that can encode a myriad of genetic circuits with applications in the fields of cell biology, therapeutics, virology, synthetic biology and biotechnology, among others. These circuits can encompass the expression of endogenous proteins, small RNAs, reporters of gene expression or cellular activity, and other innumerable reagents whose applications extend from basic discovery to medical applications. To move molecular cloning forward into an engineering discipline, with minimal trial and error and reproducible results, the parts that constitute engineered circuits need to be modular, vetted for their function and easy to share. To maximize the range of applications, circuits made from these parts need to be rapidly assembled, amenable to fast prototyping, and not bottlenecked by the delivery method to their target genomic integration sites.