PrimPol is a revolutionary brand-new enzyme obtained from the thermophilic bacteria Thermus thermophilus. PrimPol combines two distinct and complementary activities in a single thermo-stable protein: primase and polymerase. Conventional polymerases require small stretches of nucleotides (primers) annealed to a template molecule to synthesize the complementary sequence. PrimPol, on the contrary, creates its own primer sequence, thereby offering fully novel applications.
Moreover, PrimPol is able to copy both DNA and RNA. RNA reflects what genetic information is actually expressed in a cell, whereas DNA refers to the general genetic information present in every cell in the body and often only reflects a predisposition of a person to develop a disease. The development of PrimPol will help to simplify technical aspects of DNA and RNA amplification procedures and should expand the market.
PrimPol also shows a great tolerance to damaged DNA. DNA is subject to chemical modifications within the cells. Also during the processes necessary to purify the genetic material, and storage of forensic and clinical samples (e.g. formalin-fixed paraffin-embedded tissues) trigger such modifications. Chemical modifications have been shown to play an increasingly important role in several biological processes, such as aging, neurodegenerative diseases, and cancer. Therefore, there is great interest to develop methods for interrogating damaged DNA in the context of sequencing. Thus, an enzyme able to handle modified templates is of particular interest, since current amplification applications as well as second and third generation sequencing technologies are not optimized to use damaged samples.
PrimPol is also suited to be used in different second and third generation sequencing technologies due to its ability to introduce a variety of substrate nucleotides (e.g. fluorescent nucleotides) into DNA and RNA template molecules.
Finally, PrimPol has a role in multiple displacement amplification (MDA) reactions, generating primers for its subsequent use by Phi29 DNA polymerase, thus making unnecessary the use of random synthetic primers and possibly resulting in a more uniform amplification of DNA.