Philipp Holliger

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Philipp Holliger
Phil Holliger.jpg
Alma mater ETH Zurich, MRC Centre for Protein Engineering (PhD)
Scientific career
Fields Molecular biology, Synthetic biology, Xenobiology
InstitutionsMRC Centre for Protein Engineering, MRC Laboratory of Molecular Biology
Thesis Multivalent and bispecific antibody fragments from E. coli  (1994)
Doctoral advisor Sir Gregory Winter, Professor Tim Richmond
Website https://www2.mrc-lmb.cam.ac.uk/group-leaders/h-to-m/philipp-holliger/

Philipp Holliger is a Swiss molecular biologist best known for his work on xeno nucleic acids (XNAs) [1] and RNA engineering. [2] [3] Holliger is a program leader at the MRC Laboratory of Molecular Biology (MRC LMB). [4]

Contents

Background

He earned his degree in Natural Sciences (Dipl. Natwiss. ETH) from ETH Zürich, Switzerland, where he worked with Steven Benner, and his Ph.D. in Molecular Biology at the MRC Centre for Protein Engineering (CPE) in Cambridge under the mentorship of Sir Gregory Winter (CPE and MRC LMB) and Tim Richmond (ETH). [5] [6]

While in the Winter laboratory, Holliger developed a new type of bispecific antibody fragment, called a diabody and worked on elucidating the infection pathway of filamentous bacteriophages. [7] [8]

After he became an independent group leader at the MRC LMB, Holliger shifted his research focus towards synthetic biology, where he developed methods for emulsion-PCR and in vitro evolution. [9] Holliger was elected a member of EMBO in 2015. [10]

Research

XNAs

Combining nucleic acid chemistry with methods for in vitro evolution he developed, Holliger and colleagues were able to reprogram replicative DNA polymerases for the synthesis and reverse transcription of synthetic genetic polymers with entirely unnatural backbones (XNAs). This showed for the first time that synthetic alternatives to DNA could store genetic information just like DNA. [1] [11]

Further work by the Holliger lab enabled the in vitro evolution of XNA ligands (aptamers) [1] and XNA catalysts similar to RNA enzymes (known as ribozymes), termed XNAzymes [12] as well as the elaboration of simple XNA nanostructures. [13] The unnatural backbone chemistries of XNA molecules exhibit novel and useful properties. For example, unlike the natural nucleic acids, some XNAs cannot be broken down easily by the human body or are chemically much more stable. Recently, Holliger also described the synthesis and evolution of XNAs with an uncharged backbone, showing that genetic function (i.e. heredity and evolution) is possible – in contrast to previous proposals – even in the absence of a charged backbone. [14]

Origin of life

Holliger has also made contributions towards a better understanding of early steps in the origin of life. [2] [3] One scenario, termed the RNA world hypothesis, suggests that a key event in the origin of life was the emergence of an RNA molecule capable of self-replication and evolution, founding a primordial biology (lacking DNA and proteins) that relied on RNA for its main building blocks. Starting from a previously discovered ribozyme with RNA polymerase activity, Holliger and colleagues initially engineered an RNA polymerase ribozyme capable of synthesising another ribozyme [15] and subsequently RNA sequences longer than itself. [16] More recently, he described the first polymerase ribozyme that can use nucleotide triplets to copy highly structured RNA templates [17] including segments of itself.

In the course of this work, Holliger explored the properties of water ice, a simple medium likely to have been widespread on the early Earth, and found that it promotes the activity, stability and evolution of RNA polymerase ribozymes [16] and the ability of diverse pools of RNA sequences to recombine enhancing pool complexity. [18] He also discovered that the steep concentration and temperature gradients resulting from freeze-thaw cycles could be harnessed to drive ribozyme assembly and folding, acting akin to chaperones in modern biology. [19]

Related Research Articles

<span class="mw-page-title-main">DNA</span> Molecule that carries genetic information

Deoxyribonucleic acid is a polymer composed of two polynucleotide chains that coil around each other to form a double helix. The polymer carries genetic instructions for the development, functioning, growth and reproduction of all known organisms and many viruses. DNA and ribonucleic acid (RNA) are nucleic acids. Alongside proteins, lipids and complex carbohydrates (polysaccharides), nucleic acids are one of the four major types of macromolecules that are essential for all known forms of life.

<span class="mw-page-title-main">RNA</span> Family of large biological molecules

Ribonucleic acid (RNA) is a polymeric molecule that is essential for most biological functions, either by performing the function itself or by forming a template for the production of proteins. RNA and deoxyribonucleic acid (DNA) are nucleic acids. The nucleic acids constitute one of the four major macromolecules essential for all known forms of life. RNA is assembled as a chain of nucleotides. Cellular organisms use messenger RNA (mRNA) to convey genetic information that directs synthesis of specific proteins. Many viruses encode their genetic information using an RNA genome.

<span class="mw-page-title-main">RNA world</span> Hypothetical stage in the early evolutionary history of life on Earth

The RNA world is a hypothetical stage in the evolutionary history of life on Earth, in which self-replicating RNA molecules proliferated before the evolution of DNA and proteins. The term also refers to the hypothesis that posits the existence of this stage.

<span class="mw-page-title-main">Nucleobase</span> Nitrogen-containing biological compounds that form nucleosides

Nucleobases are nitrogen-containing biological compounds that form nucleosides, which, in turn, are components of nucleotides, with all of these monomers constituting the basic building blocks of nucleic acids. The ability of nucleobases to form base pairs and to stack one upon another leads directly to long-chain helical structures such as ribonucleic acid (RNA) and deoxyribonucleic acid (DNA). Five nucleobases—adenine (A), cytosine (C), guanine (G), thymine (T), and uracil (U)—are called primary or canonical. They function as the fundamental units of the genetic code, with the bases A, G, C, and T being found in DNA while A, G, C, and U are found in RNA. Thymine and uracil are distinguished by merely the presence or absence of a methyl group on the fifth carbon (C5) of these heterocyclic six-membered rings. In addition, some viruses have aminoadenine (Z) instead of adenine. It differs in having an extra amine group, creating a more stable bond to thymine.

<span class="mw-page-title-main">Peptide nucleic acid</span> Biological molecule

Peptide nucleic acid (PNA) is an artificially synthesized polymer similar to DNA or RNA.

Oligonucleotides are short DNA or RNA molecules, oligomers, that have a wide range of applications in genetic testing, research, and forensics. Commonly made in the laboratory by solid-phase chemical synthesis, these small fragments of nucleic acids can be manufactured as single-stranded molecules with any user-specified sequence, and so are vital for artificial gene synthesis, polymerase chain reaction (PCR), DNA sequencing, molecular cloning and as molecular probes. In nature, oligonucleotides are usually found as small RNA molecules that function in the regulation of gene expression, or are degradation intermediates derived from the breakdown of larger nucleic acid molecules.

<span class="mw-page-title-main">RNA polymerase</span> Enzyme that synthesizes RNA from DNA

In molecular biology, RNA polymerase, or more specifically DNA-directed/dependent RNA polymerase (DdRP), is an enzyme that catalyzes the chemical reactions that synthesize RNA from a DNA template.

<span class="mw-page-title-main">Ribozyme</span> Type of RNA molecules

Ribozymes are RNA molecules that have the ability to catalyze specific biochemical reactions, including RNA splicing in gene expression, similar to the action of protein enzymes. The 1982 discovery of ribozymes demonstrated that RNA can be both genetic material and a biological catalyst, and contributed to the RNA world hypothesis, which suggests that RNA may have been important in the evolution of prebiotic self-replicating systems.

DNA primase is an enzyme involved in the replication of DNA and is a type of RNA polymerase. Primase catalyzes the synthesis of a short RNA segment called a primer complementary to a ssDNA template. After this elongation, the RNA piece is removed by a 5' to 3' exonuclease and refilled with DNA.

Xenobiology (XB) is a subfield of synthetic biology, the study of synthesizing and manipulating biological devices and systems. The name "xenobiology" derives from the Greek word xenos, which means "stranger, alien". Xenobiology is a form of biology that is not (yet) familiar to science and is not found in nature. In practice, it describes novel biological systems and biochemistries that differ from the canonical DNA–RNA-20 amino acid system. For example, instead of DNA or RNA, XB explores nucleic acid analogues, termed xeno nucleic acid (XNA) as information carriers. It also focuses on an expanded genetic code and the incorporation of non-proteinogenic amino acids, or “xeno amino acids” into proteins.

<span class="mw-page-title-main">Aptamer</span> Oligonucleotide or peptide molecules that bind specific targets

Aptamers are short sequences of artificial DNA, RNA, XNA, or peptide that bind a specific target molecule, or family of target molecules. They exhibit a range of affinities, with variable levels of off-target binding and are sometimes classified as chemical antibodies. Aptamers and antibodies can be used in many of the same applications, but the nucleic acid-based structure of aptamers, which are mostly oligonucleotides, is very different from the amino acid-based structure of antibodies, which are proteins. This difference can make aptamers a better choice than antibodies for some purposes.

Threose nucleic acid (TNA) is an artificial genetic polymer in which the natural five-carbon ribose sugar found in RNA has been replaced by an unnatural four-carbon threose sugar. Invented by Albert Eschenmoser as part of his quest to explore the chemical etiology of RNA, TNA has become an important synthetic genetic polymer (XNA) due to its ability to efficiently base pair with complementary sequences of DNA and RNA. The main difference between TNA and DNA/RNA is their backbones. DNA and RNA have their phosphate backbones attached to the 5' carbon of the deoxyribose or ribose sugar ring, respectively. TNA, on the other hand, has it's phosphate backbone directly attached to the 3' carbon in the ring, since it does not have a 5' carbon. This modified backbone makes TNA, unlike DNA and RNA, completely refractory to nuclease digestion, making it a promising nucleic acid analog for therapeutic and diagnostic applications.

<span class="mw-page-title-main">MRC Laboratory of Molecular Biology</span> Research institute in Cambridge, England

The Medical Research Council (MRC) Laboratory of Molecular Biology (LMB) is a research institute in Cambridge, England, involved in the revolution in molecular biology which occurred in the 1950–60s. Since then it has remained a major medical research laboratory at the forefront of scientific discovery, dedicated to improving the understanding of key biological processes at atomic, molecular and cellular levels using multidisciplinary methods, with a focus on using this knowledge to address key issues in human health.

<span class="mw-page-title-main">Roger D. Kornberg</span> American biochemist and professor of structural biology

Roger David Kornberg is an American biochemist and professor of structural biology at Stanford University School of Medicine. Kornberg was awarded the Nobel Prize in Chemistry in 2006 for his studies of the process by which genetic information from DNA is copied to RNA, "the molecular basis of eukaryotic transcription."

<span class="mw-page-title-main">Nucleic acid analogue</span> Compound analogous to naturally occurring RNA and DNA

Nucleic acid analogues are compounds which are analogous to naturally occurring RNA and DNA, used in medicine and in molecular biology research. Nucleic acids are chains of nucleotides, which are composed of three parts: a phosphate backbone, a pentose sugar, either ribose or deoxyribose, and one of four nucleobases. An analogue may have any of these altered. Typically the analogue nucleobases confer, among other things, different base pairing and base stacking properties. Examples include universal bases, which can pair with all four canonical bases, and phosphate-sugar backbone analogues such as PNA, which affect the properties of the chain . Nucleic acid analogues are also called Xeno Nucleic Acid and represent one of the main pillars of xenobiology, the design of new-to-nature forms of life based on alternative biochemistries.

<span class="mw-page-title-main">Hypercycle (chemistry)</span> Cyclic sequence of self-reproducing single cycles

In chemistry, a hypercycle is an abstract model of organization of self-replicating molecules connected in a cyclic, autocatalytic manner. It was introduced in an ordinary differential equation (ODE) form by the Nobel Prize in Chemistry winner Manfred Eigen in 1971 and subsequently further extended in collaboration with Peter Schuster. It was proposed as a solution to the error threshold problem encountered during modelling of replicative molecules that hypothetically existed on the primordial Earth. As such, it explained how life on Earth could have begun using only relatively short genetic sequences, which in theory were too short to store all essential information. The hypercycle is a special case of the replicator equation. The most important properties of hypercycles are autocatalytic growth competition between cycles, once-for-ever selective behaviour, utilization of small selective advantage, rapid evolvability, increased information capacity, and selection against parasitic branches.

Numerous key discoveries in biology have emerged from studies of RNA, including seminal work in the fields of biochemistry, genetics, microbiology, molecular biology, molecular evolution and structural biology. As of 2010, 30 scientists have been awarded Nobel Prizes for experimental work that includes studies of RNA. Specific discoveries of high biological significance are discussed in this article.

<span class="mw-page-title-main">Xeno nucleic acid</span> Synthetic nucleic acid analogues

Xeno nucleic acids (XNA) are synthetic nucleic acid analogues that have a different backbone than the ribose and deoxyribose found in the nucleic acids of naturally occurring RNA and DNA.

<span class="mw-page-title-main">Hachimoji DNA</span> Synthetic DNA

Hachimoji DNA is a synthetic nucleic acid analog that uses four synthetic nucleotides in addition to the four present in the natural nucleic acids, DNA and RNA. This leads to four allowed base pairs: two unnatural base pairs formed by the synthetic nucleobases in addition to the two normal pairs. Hachimoji bases have been demonstrated in both DNA and RNA analogs, using deoxyribose and ribose respectively as the backbone sugar.

The polyelectrolyte theory of the gene proposes that for a linear genetic biopolymer dissolved in water, such as DNA, to undergo Darwinian evolution anywhere in the universe, it must be a polyelectrolyte, a polymer containing repeating ionic charges. These charges are needed to maintain the uniform physical properties needed for Darwinian evolution, regardless of the information encoded in the genetic biopolymer. DNA is such a molecule. Regardless of its nucleic acid sequence, the negative charges on its backbone dominate the physical interactions of the molecule to such a degree that it maintains uniform physical properties.

References

  1. 1 2 3 Pinheiro, Vitor B.; Taylor, Alexander I.; Cozens, Christopher; Abramov, Mikhail; Renders, Marleen; Zhang, Su; Chaput, John C.; Wengel, Jesper; Peak-Chew, Sew-Yeu; McLaughlin, Stephen H.; Herdewijn, Piet; Holliger, Philipp (20 April 2012). "Synthetic Genetic Polymers Capable of Heredity and Evolution". Science. 336 (6079): 341–344. Bibcode:2012Sci...336..341P. doi:10.1126/science.1217622. ISSN   0036-8075. PMC   3362463 . PMID   22517858.
  2. 1 2 Wochner, Aniela; Attwater, James; Coulson, Alan; Holliger, Philipp (2011-04-08). "Ribozyme-Catalyzed Transcription of an Active Ribozyme". Science. 332 (6026): 209–212. Bibcode:2011Sci...332..209W. doi:10.1126/science.1200752. ISSN   0036-8075. PMID   21474753. S2CID   39990861.
  3. 1 2 Geddes, Linda. "Earth's first life may have sprung up in ice". New Scientist. Retrieved 2021-05-16.
  4. "MRC Laboratory of Molecular Biology group leader profiles". LMB Website.
  5. "Phil Holliger - Biography". Holliger Lab Website.
  6. Holliger, Philipp (1994). Multivalent and bispecific antibody fragments from E.coli: new strategies for antibody-based diagnostics and therapeutics from bacteria. ETH Zurich (Thesis). doi:10.3929/ethz-a-001469985. hdl:20.500.11850/142158.
  7. Holliger, P.; Prospero, T.; Winter, G. (15 July 1993). ""Diabodies": small bivalent and bispecific antibody fragments". Proceedings of the National Academy of Sciences of the United States of America. 90 (14): 6444–6448. Bibcode:1993PNAS...90.6444H. doi: 10.1073/pnas.90.14.6444 . ISSN   0027-8424. PMC   46948 . PMID   8341653.
  8. Holliger, P.; Riechmann, L. (15 February 1997). "A conserved infection pathway for filamentous bacteriophages is suggested by the structure of the membrane penetration domain of the minor coat protein g3p from phage fd". Structure. 5 (2): 265–275. doi: 10.1016/s0969-2126(97)00184-6 . ISSN   0969-2126. PMID   9032075.
  9. Ghadessy, F. J.; Ong, J. L.; Holliger, P. (2001-03-27). "Directed evolution of polymerase function by compartmentalized self-replication". Proceedings of the National Academy of Sciences. 98 (8): 4552–4557. Bibcode:2001PNAS...98.4552G. doi: 10.1073/pnas.071052198 . ISSN   0027-8424. PMC   31872 . PMID   11274352.
  10. "Find people in the EMBO Communities". people.embo.org. Retrieved 2020-09-15.
  11. "Synthetic XNA molecules can evolve and store genetic information, just like DNA". Discover Magazine. Retrieved 2021-05-16.
  12. Coghlan, Andy. "Synthetic enzymes hint at life without DNA or RNA". New Scientist.
  13. Barras, Colin. "Artificial DNA folds into parcels that can survive inside us". New Scientist.
  14. Arangundy-Franklin, Sebastian; Taylor, Alexander I.; Porebski, Benjamin T.; Genna, Vito; Peak-Chew, Sew; Vaisman, Alexandra; Woodgate, Roger; Orozco, Modesto; Holliger, Philipp (June 2019). "A synthetic genetic polymer with an uncharged backbone chemistry based on alkyl phosphonate nucleic acids". Nature Chemistry. 11 (6): 533–542. Bibcode:2019NatCh..11..533A. doi:10.1038/s41557-019-0255-4. ISSN   1755-4349. PMC   6542681 . PMID   31011171.
  15. Wochner, Aniela; Attwater, James; Coulson, Alan; Holliger, Philipp (8 April 2011). "Ribozyme-Catalyzed Transcription of an Active Ribozyme". Science. 332 (6026): 209–212. Bibcode:2011Sci...332..209W. doi:10.1126/science.1200752. ISSN   0036-8075. PMID   21474753. S2CID   39990861.
  16. 1 2 Attwater, James; Wochner, Aniela; Holliger, Philipp (December 2013). "In-ice evolution of RNA polymerase ribozyme activity". Nature Chemistry. 5 (12): 1011–1018. Bibcode:2013NatCh...5.1011A. doi:10.1038/nchem.1781. ISSN   1755-4349. PMC   3920166 . PMID   24256864.
  17. Attwater, James; Raguram, Aditya; Morgunov, Alexey S; Gianni, Edoardo; Holliger, Philipp (15 May 2018). "Ribozyme-catalysed RNA synthesis using triplet building blocks". eLife. 7: e35255. doi: 10.7554/eLife.35255 . ISSN   2050-084X. PMC   6003772 . PMID   29759114. S2CID   46889517.
  18. Mutschler, Hannes; Taylor, Alexander I; Porebski, Benjamin T; Lightowlers, Alice; Houlihan, Gillian; Abramov, Mikhail; Herdewijn, Piet; Holliger, Philipp (2018-11-21). Weigel, Detlef; Muller, Ulrich (eds.). "Random-sequence genetic oligomer pools display an innate potential for ligation and recombination". eLife. 7: e43022. doi: 10.7554/eLife.43022 . ISSN   2050-084X. PMC   6289569 . PMID   30461419.
  19. Mutschler, Hannes; Wochner, Aniela; Holliger, Philipp (June 2015). "Freeze–thaw cycles as drivers of complex ribozyme assembly". Nature Chemistry. 7 (6): 502–508. Bibcode:2015NatCh...7..502M. doi:10.1038/nchem.2251. ISSN   1755-4349. PMC   4495579 . PMID   25991529.