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10:29
Inducing Cre-lox Recombination in Mouse Cerebral Cortex Through In Utero Electroporation
Published on: November 17, 2017
Using the Cre-lox system to randomize target gene expression states and generate diverse phenotypes
Bradley Niesner1, Narendra Maheshri
1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Biotechnology and Bioengineering
|June 5, 2013
Summary
This study introduces a novel Cre-loxP recombination method to randomly alter gene expression in yeast. This technique simplifies the engineering of complex cellular phenotypes and gene function studies.
Area of Science:
- Synthetic Biology
- Molecular Biology
- Yeast Genetics
Background:
- Engineering complex cellular phenotypes and understanding gene function requires modifying multiple genes.
- Traditional methods for multigene modification are laborious and involve sequential genetic alterations.
Purpose of the Study:
- To develop a strategy for randomizing the expression states of multiple genes in Saccharomyces cerevisiae.
- To enable efficient engineering of multigenic cellular phenotypes and facilitate gene function discovery.
Main Methods:
- Utilized Cre-loxP recombination by inserting promoters flanked by inverted loxP sites upstream of target genes.
- Randomized gene expression to ON, OFF, or four distinct intermediate states.
- Combined the strategy with yeast mating to introduce native gene regulation.
Main Results:
- Demonstrated independent randomization of at least 6 genes.
- Proposed that using orthogonal loxP sites could increase the number of independently randomized genes to at least 15.
- Successfully probed the role of four base excision repair enzymes in methyl methanesulfonate (MMS) tolerance.
Conclusions:
- The developed Cre-loxP system provides a versatile tool for randomizing gene expression in yeast.
- This strategy is applicable to both heterologous and endogenous genes, with potential for metabolic engineering.
- The method's adaptability across organisms suggests broad future applications in genetic engineering.
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