Table of Contents
- 1 Why is Rosa26 used?
- 2 What chromosome is in the Rosa26 locus?
- 3 Where is ROSA26 locus?
- 4 What is the AAVS1 locus?
- 5 How does CRE ERT2 work?
- 6 Why would a scientist want to Knockin a gene?
- 7 Where is AAVS1?
- 8 What is safe harbor locus?
- 9 What is the ROSA26 locus and why is it important?
- 10 Why is ROSA26 used for Transgene expression?
Why is Rosa26 used?
The Rosa26 locus is a useful place for inserting a gene, The location of the insertion is known — not random — and it allows scientists to study a gene without affecting the function of other genes. Knock-in models using this locus offer greater accuracy and reproducibility of results.
What chromosome is in the Rosa26 locus?
chromosome 6
The Rosa26 locus on chromosome 6 is frequently used for the integration of transgene constructs to achieve ubiquitous or conditional gene expression in mice. The Rosa26 transcript is spliced into three exons and ubiquitously expressed in all cell types and developmental stages, but not translated to a protein [1].
What is Rosa26 cre?
These R26CreER mutant mice have a tamoxifen-inducible Cre-mediated recombination system driven by the endogenous mouse Gt(ROSA)26Sor promoter. When crossed with a strain containing a loxP site-flanked sequence of interest, this mutant is useful for generating tamoxifen-induced, Cre-mediated targeted deletions.
Where is ROSA26 locus?
mouse chromosome 6
Known in scientific circles as Gt(ROSA)26Sor, the Rosa26 locus is a specific genetic site that is located on mouse chromosome 6, and is considered to be one of the ideal locations where knock-ins of interest can be targeted.
What is the AAVS1 locus?
The adeno-associated virus site 1 (AAVS1) locus in the human genome is a strong candidate for gene therapy by insertion of an exogenous gene into the locus. The AAVS1 locus includes the coding region for myosin binding subunit 85 (MBS85).
Where is ROSA26?
Known in scientific circles as Gt(ROSA)26Sor, the Rosa26 locus is a specific genetic site that is located on mouse chromosome 6, and is considered to be one of the ideal locations where knock-ins of interest can be targeted.
How does CRE ERT2 work?
The Cre-ERT2 fusion protein (Cre-ERT2) consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor; which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with …
Why would a scientist want to Knockin a gene?
Knocking in genes allows scientists to study the effects of specific gene variants, to use reporter genes like green fluorescent protein to track gene products in time and space, to probe genome regulation, and ultimately, to repair disease-causing genes.
What are Knockin mice?
A Knockin mouse defines an animal model in which a gene sequence of interest is altered by one-for-one substitution with a transgene, or by adding gene sequences that are not found within the locus. The insertion of a transgene is typically done in specific loci.
Where is AAVS1?
chromosome 19
Adeno-associated virus integration site 1 is a viral integration site that in humans is encoded by the AAVS1 gene located on chromosome 19.
What is safe harbor locus?
The AAVS1 locus, in intron 1 of PPP1R12C (protein phosphatase 1 regulatory subunit 12C), is known as a “safe harbor” because its disruption does not have adverse effects on the cell, and robust transcription can be used to maintain the expression of an exogenously inserted gene5,6,7.
How is ROSA26 expressed in mice?
It is moderately expressed with levels varying between different tissues in pre and post natal life. Currently, Rosa26 is a safe harbor locus broadly used for both constitutive and conditional gene expression in mice. It is proven safe in mice because the transgene is adequately expressed without perturbing endogenous gene structure or function.
What is the ROSA26 locus and why is it important?
The Rosa26 locus is one of most stable places to target single transgenes. In most cases, when Rosa26 is used, there’s a high rate of success when it comes to the correct insertion of the transgene and the facilitation of a stable, single-copy expression.
Why is ROSA26 used for Transgene expression?
In most cases, when Rosa26 is used, there’s a high rate of success when it comes to the correct insertion of the transgene and the facilitation of a stable, single-copy expression. Rosa26 also allows for a smaller number of mice because a targeted gene knock-in strategy is being used.
Is it possible to use ROSA26 in humans?
While studying gene insertions in mice could help future human genetics research, the use of Rosa26 in humans is not yet possible. Unlike the location in mice, Rosa26 in humans is close to critical genes. Gene editing the Rosa26 locus in humans could, therefore, disrupt the function of these other genes.