Abstract
Covering: up to the end of 2015 While Nature continues to deliver a myriad of potent and structurally diverse biologically active small molecules, the cellular targets and modes of action of these natural products are rarely identified, significantly hindering their development as new chemotherapeutic agents. This article provides an introductory tutorial on the use of T7 phage display as a tool to rapidly identify the cellular targets of natural products and is aimed specifically at natural products chemists who may have only limited experience in molecular biology. A brief overview of T7 phage display is provided, including its strengths, weaknesses, and the type of problems that can and cannot be tackled with this technology. Affinity probe construction is reviewed, including linker design and natural product derivatisation strategies. A detailed description of the T7 phage biopanning procedure is provided, with valuable tips for optimising each step in the process, as well as advice for identifying and avoiding the most commonly encountered challenges and pitfalls along the way. Finally, a brief discussion is provided on techniques for validating the cellular targets identified using T7 phage display.
| Language | English |
|---|---|
| Pages | 626-636 |
| Number of pages | 11 |
| Journal | Natural Product Reports |
| Volume | 33 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 1 May 2016 |
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Identifying the cellular targets of natural products using T7 phage display. / Piggott, Andrew M.; Karuso, Peter.
In: Natural Product Reports, Vol. 33, No. 5, 01.05.2016, p. 626-636.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Identifying the cellular targets of natural products using T7 phage display
AU - Piggott, Andrew M.
AU - Karuso, Peter
PY - 2016/5/1
Y1 - 2016/5/1
N2 - Covering: up to the end of 2015 While Nature continues to deliver a myriad of potent and structurally diverse biologically active small molecules, the cellular targets and modes of action of these natural products are rarely identified, significantly hindering their development as new chemotherapeutic agents. This article provides an introductory tutorial on the use of T7 phage display as a tool to rapidly identify the cellular targets of natural products and is aimed specifically at natural products chemists who may have only limited experience in molecular biology. A brief overview of T7 phage display is provided, including its strengths, weaknesses, and the type of problems that can and cannot be tackled with this technology. Affinity probe construction is reviewed, including linker design and natural product derivatisation strategies. A detailed description of the T7 phage biopanning procedure is provided, with valuable tips for optimising each step in the process, as well as advice for identifying and avoiding the most commonly encountered challenges and pitfalls along the way. Finally, a brief discussion is provided on techniques for validating the cellular targets identified using T7 phage display.
AB - Covering: up to the end of 2015 While Nature continues to deliver a myriad of potent and structurally diverse biologically active small molecules, the cellular targets and modes of action of these natural products are rarely identified, significantly hindering their development as new chemotherapeutic agents. This article provides an introductory tutorial on the use of T7 phage display as a tool to rapidly identify the cellular targets of natural products and is aimed specifically at natural products chemists who may have only limited experience in molecular biology. A brief overview of T7 phage display is provided, including its strengths, weaknesses, and the type of problems that can and cannot be tackled with this technology. Affinity probe construction is reviewed, including linker design and natural product derivatisation strategies. A detailed description of the T7 phage biopanning procedure is provided, with valuable tips for optimising each step in the process, as well as advice for identifying and avoiding the most commonly encountered challenges and pitfalls along the way. Finally, a brief discussion is provided on techniques for validating the cellular targets identified using T7 phage display.
UR - http://www.scopus.com/inward/record.url?scp=84973500638&partnerID=8YFLogxK
UR - http://purl.org/au-research/grants/arc/DP130103281
UR - http://purl.org/au-research/grants/arc/FT130100142
U2 - 10.1039/c5np00128e
DO - 10.1039/c5np00128e
M3 - Article
VL - 33
SP - 626
EP - 636
JO - Natural Product Reports
T2 - Natural Product Reports
JF - Natural Product Reports
SN - 0265-0568
IS - 5
ER -