Latest research - September 2023
Irish Examiner article featuring Dr Harrison's research
Dr Harrison gives evidence to Oireachtas committee on gene editing
https://oireachtas.ie/en/oireachtas-tv/video-archive/committees/5420/… (0:41:09 onwards)
GENE EDITING FOR RESEARCH AND THERAPY
The research focus of the Harrison Lab is the development of CRISPR gene, base and prime editing to study and potentially treat genetic disorders such as cystic fibrosis (CF).
What is CRISPR gene editing?
CRISPR gene editing is a Nobel Prize winning technique (https://www.nobelprize.org/prizes/chemistry/2020/summary/) that enables the sequence of DNA of a living cell to be precisely changed, and has at least three major biomedical applications:
- First, this ability to precisely change a DNA sequence within a cell allows researchers to learn how particular DNA sequences control the normal structure and functions of individual cells.
- Second, it enables us to learn of how certain mutations in the DNA sequence, either acquired or inherited, can cause more than 100 different types of cancer or over 7,000 genetic disorders.
- Third, gene editing could be developed as the basis for a potential treatments for many of these diseases, such as cystic fibrosis (CF), sickle-cell anaemia and cystinosis.
How are we using CRISPR to understand Cystic Fibrosis?
We are currently working on three new approaches Base Editing, HITI and Prime editing as outlined in recent publications
https://pubmed.ncbi.nlm.nih.gov/35313109/ - CRISPR for respiratory medicine (2022)
https://pubmed.ncbi.nlm.nih.gov/34103250/ - CF gene editing (2022)
https://pubmed.ncbi.nlm.nih.gov/34031549/ - Prime Editing (2021)
https://pubmed.ncbi.nlm.nih.gov/31117296/ - Base Editing and Disease (2019)
https://pubmed.ncbi.nlm.nih.gov/29233638/ - Gene Editing and Stem Cells (2018)
This work was built on some of our earlier studies using novel CRISPR approaches to correct other CF causing variants
https://pubmed.ncbi.nlm.nih.gov/28863137/ - targeted excision of intronic variants (2017)
https://pubmed.ncbi.nlm.nih.gov/27557525/ - improved HDR with CRISPR (2016)
In 2012, we published our breakthrough paper on the use of ZFN to correct the most common CF mutation known as F508del.
https://pubmed.ncbi.nlm.nih.gov/23514673/ - first gene editing study to correct a CF mutation
How are we using CRISPR to understand other diseases?
In the last year, we have published on use of CRISPR and development as a therapeutic tool with several collaborators in Liverpool, Auckland and beyond.
https://pubmed.ncbi.nlm.nih.gov/35011573/ - CRISPR in vivo for Cystinosis (2022)
https://pubmed.ncbi.nlm.nih.gov/33671852/ - CRISPR to study gene regulation (2021)
https://www.karger.com/Article/Pdf/511678/ - ATMP policy discussion (2021)
https://pubmed.ncbi.nlm.nih.gov/32198276/ - Cystinosis iPSc model (2020)
Our current work is supported by grants from CF Trust (UK), CF Foundation (USA) and Science Foundation Ireland.
Research Interests2020 onwards - Prime Editing, ABE and HITI for Cystic Fibrosis
The current focus of the Harrison Lab is the development of gene, base and prime editing to repair disease-causing mutations in the human genome funded by CFF foundation (USA) and CF Trust (UK)
2017 to 2019 - CRISPR NHEJ and HITI for Cystic Fibrosis and Atopic Dermatitis
The focus was on NHEJ based methods. A published study showed effective excision of CF-causing deep intronic mutations (Sanz et al., 2017), but focus moved to use of HITI to correct multiple mutations with a single donor and Cas9/gRNA combination. This work was supported by grants from CF Foundation Therapeutics (USA) and CF Trust (UK). The lab was also a member of the CF Trust's first Strategic Research Centre for Gene Editing during this period, and colaboration with Carlos Farinha in Lisbon to create novel cell lines to study rare CF mutations (part funded by CF Foundation) using gene and base editing
2013 to 2016 - CRISPR HDR for Cystic Fibrosis
In 2013, the lab starting using the CRISPR Cas9/gRNA system for gene editing to correct the F508del mutation by HDR (Hollywood et al., 2016) with grants from CF Foundation Therapeutics (USA) and CF Trust (UK). The lab has funded collaborative projects with Alan Davidson and Jennifer Hollywood in Auckland (funded by Cystinosis Ireland and Health Research Board), and Isabelle Sermet in Paris (funded by HRB).
2005 to 2012 - Application of ZFN HDR to cystic fibrosis
Since 2005, my lab has been working on the production of ZFNs to target the most common gene defect in patients with cystic fibrosis. This work was started by a PhD student, Rowan Flynn, and is currently under further investigation by Ciaran Lee and Jennifer Hollywoord. Using an in vitro system, we developed ZFNs which can target and correct the F508del mutation in the cftr gene http://online.liebertpub.com/doi/abs/10.1089/biores.2012.0218
2008 to 2012 - Application of ZFN HDR to cystinosis
Cystinosis is a rare genetic disorder affecting less than 10 individuals in Ireland. The disease which is a multisystem disorder invariably leads to kidney failure. The gene defect was characterised in 1999, and Dr. Harrison and Dr. Scallan were recently been awarded funding from Cystinosis Ireland (www.cystinosis.ie) and the HRB (www.hrb.ie), as well as the Cystinosis Research Foundation (www.natalieswish.org) to study the applicability of ZFN HDR technology, virus vector delivery and Tale nuclease gene repair. This work has been conducted by Katrin Kaschig and Ciaran Lee.
2001 to 2007 - Molecular Physiology
Between 2001 and 2007, the Department hosted five 10-day residential workshop to provide training for physiologists in molecular biological techniques (Harrison, 2004). The workshop was sponsored by the Physiological Society and the Wellcome Trust, and offered training in experimental procedures in molecular physiology, including the handling of DNA and RNA, sub-cloning, restriction enzyme digests, the use of siRNA technology, western blots, RT-PCR, site-directed mutagenesis and transfection techniques. During this time over 80 physiologists at PhD or post-doctoral level attended these workshops. Further details of other courses organised by the physiological society can be found at www.physoc.org. In 2010, the Department hosted two shorter workshops for physiologists, again sponsored by the Physiological Society.
|Start Date||End Date||Award|
|Gene Editing of CFTR.||Foreign Research Institute||01-OCT-21||30-SEP-24||€160,542.00|
|Project Title: 25,000 non-drug alleles fixed with 15 (or less) CRISPR PRIME pegRNAs||Foundation Funded Research||01-JUL-20||30-JUN-24||€247,856.00|
|Temporal-spatial CRISPR Base Editing to Identify Target Cells for CFTR Repair||Foreign Research Institute||01-JUL-21||30-JUN-24||€156,386.00|
|Cystic Fibrosis Trust||Foundation Funded Research||01-MAY-16||31-MAR-20||€152,003.00|
|Splicing mutations||Foreign Research Institute||17-NOV-14||31-JAN-20||€59,963.00|
|Permanent correction of 80% of Disease causing Mutations in Human CF cells.||Foreign Research Institute||01-OCT-13||31-JAN-20||€62,458.00|
|Down's Symdrome Study||Foreign Research Institute||22-FEB-18||31-DEC-19||€1,000.00|
|Isogenic Cell Models to Chareterise Disease Siganutures of Seven CF-Causing Mutations and HITI-Gene Editing to Correct Them.||Foundation Funded Research||01-DEC-17||30-NOV-19||€46,900.00|
|Isogenic models to study CF disease signatures: HITI gene edit to fix them||Foundation Funded Research||01-OCT-17||30-SEP-19||€117,945.00|
|Cystic Fibrosis- Gene Editing of Non-Druggable CFTR Mutations.||Irish Research Council||01-FEB-17||30-JUN-18||€2,425.00|
|Cystic Fibrosis Foundation Therapeutics CRISPR Knock-Out of Class 1 CF Splicing Mutations.||Foreign Research Institute||01-OCT-14||31-MAY-18||€162,902.00|
|Modelling cystinosis with human stem cells and the therapeutic potential of aspartic acid||Health Research Board||01-NOV-14||30-OCT-17||€295,000.00|
|Science Foundation of Ireland||01-FEB-16||31-JUL-16||€3,000.00|
Peer Reviewed Journals
|(2021)||'CRISPR Deletion of a SVA Retrotransposon Demonstrates Function as a cis-Regulatory Element at the TRPV1/TRPV3 Intergenic Region' |
Price E;Gianfrancesco O;Harrison PT;Frank B;Bubb VJ;Quinn JP; (2021) 'CRISPR Deletion of a SVA Retrotransposon Demonstrates Function as a cis-Regulatory Element at the TRPV1/TRPV3 Intergenic Region'. International Journal of Molecular Sciences, 22 (4) [DOI] [Details]
|(2021)||'Prime editing - an update on the field' |
Scholefield J;Harrison PT; (2021) 'Prime editing - an update on the field'. Gene Therapy, 28 (7-8) [DOI] [Details]
|(2021)||'Comparison of Cas9 and Cas12a CRISPR editing methods to correct the W1282X-CFTR mutation' |
Santos L;Mention K;Cavusoglu-Doran K;Sanz DJ;Bacalhau M;Lopes-Pacheco M;Harrison PT;Farinha CM; (2021) 'Comparison of Cas9 and Cas12a CRISPR editing methods to correct the W1282X-CFTR mutation'. Journal of Cystic Fibrosis, [DOI] [Details]
|(2020)||'Polycystic kidney disease ryanodine receptor domain (Pkdrr) proteins in oomycetes' |
Zheng L.;Prestwich B.D.;Harrison P.T.;Mackrill J.J. (2020) 'Polycystic kidney disease ryanodine receptor domain (Pkdrr) proteins in oomycetes'. Journal of Pathogens, 9 (7):1-19 [DOI] [Details]
|(2019)||'Minigene Assay to Evaluate CRISPR/Cas9-based Excision of Intronic Mutations that Cause Aberrant Splicing in Human Cells' |
Sanz, DJ;Harrison, PT (2019) 'Minigene Assay to Evaluate CRISPR/Cas9-based Excision of Intronic Mutations that Cause Aberrant Splicing in Human Cells'. Bio-Protocol, 9 [DOI] [Details]
|(2019)||'Gene and Base Editing as a Therapeutic Option for Cystic Fibrosis-Learning from Other Diseases' |
Mention K;Santos L;Harrison PT; (2019) 'Gene and Base Editing as a Therapeutic Option for Cystic Fibrosis-Learning from Other Diseases'. Genes, 10 (5) [DOI] [Details]
|(2018)||'The Regulation of Monoamine Oxidase A Gene Expression by Distinct Variable Number Tandem Repeats' |
Manca, M;Pessoa, V;Lopez, AI;Harrison, PT;Miyajima, F;Sharp, H;Pickles, A;Hill, J;Murgatroyd, C;Bubb, VJ;Quinn, JP (2018) 'The Regulation of Monoamine Oxidase A Gene Expression by Distinct Variable Number Tandem Repeats'. Journal of Molecular Neuroscience, 64 :459-470 [DOI] [Details]
|(2017)||'Cas9/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA' |
Sanz, DJ;Hollywood, JA;Scallan, MF;Harrison, PT (2017) 'Cas9/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA'. Plos One, 12 [DOI] [Details]
|(2018)||'Gene Editing of Stem Cells to Model and Treat Disease' |
Hollywood, JA;Sanz, DJ;Davidson, AJ;Harrison, PT (2018) 'Gene Editing of Stem Cells to Model and Treat Disease'. Current Stem Cell Reports, 4 :253-263 [DOI] [Details]
|(2018)||'A beginner's guide to gene editing' |
Harrison, Patrick T.; Hart, Stephen (2018) 'A beginner's guide to gene editing'. Experimental Physiology, [DOI] [Full Text] [Details]
|(2017)||'Double TALEN-edited T-cells kick B-ALL into touch' |
Harrison PT (2017) 'Double TALEN-edited T-cells kick B-ALL into touch'. Gene Therapy, 24 (3) [Details]
|(2017)||'Cas9/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA' |
Sanz DJ, Hollywood JA, Scallan MF, Harrison PT (2017) 'Cas9/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA'. Plos One, 12 (9) [Details]
|(2017)||'Genetic therapies for cystic fibrosis lung disease' |
Hart, SL;Harrison, PT (2017) 'Genetic therapies for cystic fibrosis lung disease'. Current Opinion In Pharmacology, 34 :119-124 [DOI] [Details]
|(2016)||'Analysis of gene repair tracts from Cas9/gRNA double-stranded breaks in the human CFTR gene' |
Hollywood, JA,Lee, CM,Scallan, MF,Harrison, PT (2016) 'Analysis of gene repair tracts from Cas9/gRNA double-stranded breaks in the human CFTR gene'. Scientific Reports, 6 [DOI] [Details]
|(2016)||'Impact of gene editing on the study of cystic fibrosis' |
Harrison PT;Sanz DJ;Hollywood JA; (2016) 'Impact of gene editing on the study of cystic fibrosis'. Human Genetics, 135 (9) [DOI] [Details]
|(2016)||'Genetic medicines for CF: Hype versus reality' |
Alton EW;Boyd AC;Davies JC;Gill DR;Griesenbach U;Harrison PT;Henig N;Higgins T;Hyde SC;Innes JA;Korman MS; (2016) 'Genetic medicines for CF: Hype versus reality'. Pediatric Pulmonology, 51 (S44) [DOI] [Details]
|(2012)||'Correction of the ΔF508 mutation in the CFTR gene by zinc finger nuclease homology-directed repair' |
Lee, CM, Flynn, R, Hollywood, JA, Scallan, MF and Harrison, PT (2012) 'Correction of the ΔF508 mutation in the CFTR gene by zinc finger nuclease homology-directed repair'. BioResearch Open Access, 1 (2):99-108 [DOI] [Details]
|(2017)||Double TALEN-edited T-cells kick B-ALL into touch. |
Harrison, PT (2017) Double TALEN-edited T-cells kick B-ALL into touch. LONDON: Editorship [DOI] [Details]
|(2019)||Gene and Base Editing as a Therapeutic Option for Cystic FibrosisLearning from Other Diseases. |
Mention, K;Santos, L;Harrison, PT (2019) Gene and Base Editing as a Therapeutic Option for Cystic FibrosisLearning from Other Diseases. BASEL: Reviews [DOI] [Details]
|(2018)||Gene editing & stem cells. |
Harrison, PT;Hoppe, N;Martin, U (2018) Gene editing & stem cells. AMSTERDAM: Reviews [DOI] [Details]
|(2016)||Impact of gene editing on the study of cystic fibrosis. |
Harrison, PT;Sanz, DJ;Hollywood, JA (2016) Impact of gene editing on the study of cystic fibrosis. NEW YORK: Reviews [DOI] [Details]
|(2008)||Viral vectors in cancer immunotherapy: Which vector for which strategy?. |
Collins, SA;Guinn, BA;Harrison, PT;Scallan, MF;O'Sullivan, GC;Tangney, M (2008) Viral vectors in cancer immunotherapy: Which vector for which strategy?. SHARJAH: Reviews [Details]
Teaching InterestsTeaching interest include CRISPR gene editing and gene therapy, molecular physiology, and introductory physiology.
|Graduation Year||Student Name||Institution||Degree Type||Thesis Title|
|2022||Lúcia Santos||Technical University of Lisbon||PHD|
|2020||Karen Mention||NUI (UCC)||PHD||CRISPR Editing of CFTR|
|2013||Jennifer Hollywood||PHD||Cystic fibrosis gene repair : correction of ΔF508 using ZFN and CRISPR/Cas9 guide RNA gene editing tools|
|2013||Katrin Kaschig||PHD||Zinc finger nuclease gene repair as a treatment for cystinosis|
|2013||Ana Marie Landin||PHD||Molecular regulation of B cell deficiency with age : a study of E47 and Pax-5|
|2011||Sarah Collins||PHD||Adeno-Associated virus vectors for cancer gene therapy|
|2010||Ciaran Lee||UCC||PHD||Cystic fibrosis gene repair : development of zinc finger nucleases for homology directed repair of the CFTR|
|2007||Rowan Flynn||UCC||PHD||1 THESIS Cystic fibrosis gene repair : design and evaluation of zinc finger nucleases for homology-directed r|
|2006||David B O'Sullivan||UCC||PHD||Establishment of growth/differentiation factor 5-expressing cells for transplantation in a model of Parkinson's disease|
|2001||Melanie McCarthy-Troke||University of Glasgow||PHD|
|2001||Pavel Mistrik||University of Glasgow||PHD|