BioBricks++: Simplifying Assembly of Standard DNA Components

Abstract

Construction of complex biological systems can require assembling many modules together. However, existing assembly schemes are lacking in generality, ease of use, or power to perform some desirable operations. Currently, biological modules are most easily specified and manipulated as DNA sequences. A general system, called BioBricks++, for assembling and manipulating DNA modules is proposed. BioBricks++ was inspired by the BioBricks assembly scheme but provides for more possible module operations.

BioBricks++ uses commercially available restriction enzymes and standard biological techniques for assembling modules. The key to the method is in the specification of the standard DNA module. Modules are packaged with a standard prefix and suffix DNA sequence containing several restriction enzyme sites, which are used for different module operations.

The following operations can be performed on all BioBricks++ modules. The most fundamental operation is the arbitrary assembly of any two modules. In addition, the assembly of the two modules can be made seamless, with no extra intervening sequence inserted between the modules. Modules can also be easily reversed with no extra bases added during the operation. Another useful capability is being able to remove bases from either end of a module, allowing for operations such as protein fusions or addition of tags.