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The use of restriction enzymes is no option for cloning if functional nucleic acids contain the recognition sites for enzymes used in standard restriction cloning, as described in RFC 10, or other suitable restriction enzymes. Although techniques using Type IIs restriction enzymes like Golden Gate assembly do not leave scars after cloning, a Type IIs recognition site within the functional RNA sequence may greatly reduce the efficiency and fidelity of a Golden Gate based cloning attempt. Therefore, the method of choice for reliable assembly of the sequences into a standardized vector has to be based on homology at the interfaces of the parts to be fused. As the DNA templates for functional RNA are commonly synthesized de novo, the addition of overhangs upstream and downstream the functional sequence does not appear to be challenging. Equally those extensions can be added during PCR with primer overhangs.

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The term “functional RNA” covers a wide range of noncoding natural and synthetic RNA. These RNAs do not need to be translated into protein and are able to fulfil their function either by simple Watson-Crick basepairing interactions or by a more complex formation of secondary structures. Noteworthy classes of functional RNA relevant to the synthetic biologist include:

The term “functional RNA” covers a wide range of noncoding natural and synthetic RNA. These RNAs do not need to be translated into protein and are able to fulfil their function either by simple Watson-Crick basepairing interactions or by a more complex formation of secondary structures. Noteworthy classes of functional RNA relevant to the synthetic biologist include: