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#conditional knockout

2 public questions tagged with this topic.

Gene Tar getting

The ‘floxed’ gene refers to a gene:

Floxed allele describes engineered genomic locus where essential region is flanked by loxP sequences, derived from phrase flanked by loxP. Construction inserts 34 base pair loxP sites into introns bracketing target exon via homologous recombination in embryonic stem cells, without disrupting transcription. Without Cre, floxed gene expresses normally. Upon crossing to Cre driver, recombination deletes floxed segment in desired tissue. Floxed terminology distinguishes conditional ready allele from constitutive knockout, enabling temporal control, lineage tracing and rescue experiments in mouse genetics.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Gene Tar getting

In conditional knockout using Cre, gene excision happens when:

Conditional knockout overcomes embryonic lethality by floxing essential exons with loxP sites placed in same orientation flanking critical region intronically, preserving normal splicing without Cre. Excision occurs only upon Cre expression which recombines directly repeated loxP sites, deleting intervening exon and creating frameshift or null allele. Tissue specific or tamoxifen inducible Cre drivers then restrict deletion temporally and spatially. Requirement of same direction loxP ensures deletion rather than inversion, producing irreversible loss of function in selected cells only in vivo efficiently.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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