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Updated: Jun 5, 2026
13:51
The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
BAC modification through serial or simultaneous use of CRE/Lox technology
Mark Parrish1, Jay Unruh, Robb Krumlauf
1Krumlauf Laboratory, Stowers Institute for Medical Research, 1000 50th Street, Kansas City, MO 64110, USA.
Journal of Biomedicine & Biotechnology
|January 4, 2011
Summary
This study explores Lox variant sites for modifying bacterial artificial chromosomes (BACs). Researchers demonstrate specific serial and simultaneous BAC manipulations, enabling complex genomic engineering in mice.
Area of Science:
- Genomics
- Molecular Biology
- Biotechnology
Background:
- Bacterial artificial chromosomes (BACs) are essential for large DNA inserts in mouse genomics.
- Conventional molecular techniques are limited for modifying large BAC constructs.
Purpose of the Study:
- To investigate the use of Lox variant sites for serial and simultaneous BAC manipulation.
- To enhance methods for engineering large DNA constructs.
Main Methods:
- Utilized Lox variant sites, including spacer mutants and inverted repeat variants.
- Applied recombineering and Cre/Lox methodologies for BAC modification.
- Performed serial and simultaneous genetic alterations on BACs.
Main Results:
- Demonstrated high specificity of Lox spacer mutants.
- Showed that inverted repeat variants reduce Cre binding affinity while retaining functionality.
- Successfully generated a mouse HoxB BAC with four reporter gene insertions.
- Achieved specific, simultaneous deletions using combinations of Lox variants.
Conclusions:
- Lox variant sites offer precise tools for complex BAC engineering.
- These methods expand the repertoire for BAC and genome manipulation.
- Facilitates advanced mouse genomic analyses and genetic engineering.
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