Limits...
RMRP is a non-coding RNA essential for early murine development.

Rosenbluh J, Nijhawan D, Chen Z, Wong KK, Masutomi K, Hahn WC - PLoS ONE (2011)

Bottom Line: RMRP mutations that lead to decreased RMRP levels are found in the pleiotropic syndrome Cartilage Hair Hypoplasia.To assess the effects of deleting RMRP, we engineered a targeting vector that contains loxP sequences flanking RMRP and created hemizygous mice harboring this engineered allele (RMRP conditional).We found that insertion of this cassette suppressed RMRP expression, and we failed to obtain viable mice homozygous for the RMRP conditional allele.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
RMRP is a non-coding RNA that is ubiquitously expressed in both humans and mice. RMRP mutations that lead to decreased RMRP levels are found in the pleiotropic syndrome Cartilage Hair Hypoplasia. To assess the effects of deleting RMRP, we engineered a targeting vector that contains loxP sequences flanking RMRP and created hemizygous mice harboring this engineered allele (RMRP conditional). We found that insertion of this cassette suppressed RMRP expression, and we failed to obtain viable mice homozygous for the RMRP conditional allele. Furthermore, we were unable to obtain viable homozygous RMRP mice, indicating that RMRP is essential for early embryonic development.

Show MeSH

Related in: MedlinePlus

Genes near RMRP are not essential for cellular viability.MEFs from E13.5 mice of either A. WT or B. RMRP+/− were transfected with siRNAs targeting Ccdc107 or E130. Three days later RNA was extracted from the cells and qRT-PCR was preformed using primers for RMRP, Ccdc107 or E130. C. The same cells as in A and B were plated (5000 cells/well) in a 96 well plate and 7 days post transfection cell number was assessed by Cell titer glow. Error bars represent SD of three replicas.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3198473&req=5

pone-0026270-g004: Genes near RMRP are not essential for cellular viability.MEFs from E13.5 mice of either A. WT or B. RMRP+/− were transfected with siRNAs targeting Ccdc107 or E130. Three days later RNA was extracted from the cells and qRT-PCR was preformed using primers for RMRP, Ccdc107 or E130. C. The same cells as in A and B were plated (5000 cells/well) in a 96 well plate and 7 days post transfection cell number was assessed by Cell titer glow. Error bars represent SD of three replicas.

Mentions: Based on these observations, we tested whether Ccdc107 or E130 are essential for survival and thus may also contribute to the observed embryonic lethal phenotype. We used MEFs from E13.5 WT or RMRP+/− mice. Using RNAi, we reduced the expression of these genes to 5–20% of levels found in cells transfected with a control siRNA (Figure 4A and B). Seven days post transfection the cells were tested for viability, by monitoring the ATP content of the cells (Figure 4C). We failed to observe any alteration in cell proliferation/viability after suppression of either Ccdc107 or E130 indicating that these genes are not essential. However, we cannot eliminate the possibility that the residual low levels of Ccdc107 or E130 (5–20% of control) are enough to sustain viability and when completely deleted will lead to embryonic lethality. Due to the very close proximity between these genes it is very difficult to target RMRP without disrupting Ccdc107 and E130 expression and further attempts to target RMRP should take this into consideration.


RMRP is a non-coding RNA essential for early murine development.

Rosenbluh J, Nijhawan D, Chen Z, Wong KK, Masutomi K, Hahn WC - PLoS ONE (2011)

Genes near RMRP are not essential for cellular viability.MEFs from E13.5 mice of either A. WT or B. RMRP+/− were transfected with siRNAs targeting Ccdc107 or E130. Three days later RNA was extracted from the cells and qRT-PCR was preformed using primers for RMRP, Ccdc107 or E130. C. The same cells as in A and B were plated (5000 cells/well) in a 96 well plate and 7 days post transfection cell number was assessed by Cell titer glow. Error bars represent SD of three replicas.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198473&req=5

pone-0026270-g004: Genes near RMRP are not essential for cellular viability.MEFs from E13.5 mice of either A. WT or B. RMRP+/− were transfected with siRNAs targeting Ccdc107 or E130. Three days later RNA was extracted from the cells and qRT-PCR was preformed using primers for RMRP, Ccdc107 or E130. C. The same cells as in A and B were plated (5000 cells/well) in a 96 well plate and 7 days post transfection cell number was assessed by Cell titer glow. Error bars represent SD of three replicas.
Mentions: Based on these observations, we tested whether Ccdc107 or E130 are essential for survival and thus may also contribute to the observed embryonic lethal phenotype. We used MEFs from E13.5 WT or RMRP+/− mice. Using RNAi, we reduced the expression of these genes to 5–20% of levels found in cells transfected with a control siRNA (Figure 4A and B). Seven days post transfection the cells were tested for viability, by monitoring the ATP content of the cells (Figure 4C). We failed to observe any alteration in cell proliferation/viability after suppression of either Ccdc107 or E130 indicating that these genes are not essential. However, we cannot eliminate the possibility that the residual low levels of Ccdc107 or E130 (5–20% of control) are enough to sustain viability and when completely deleted will lead to embryonic lethality. Due to the very close proximity between these genes it is very difficult to target RMRP without disrupting Ccdc107 and E130 expression and further attempts to target RMRP should take this into consideration.

Bottom Line: RMRP mutations that lead to decreased RMRP levels are found in the pleiotropic syndrome Cartilage Hair Hypoplasia.To assess the effects of deleting RMRP, we engineered a targeting vector that contains loxP sequences flanking RMRP and created hemizygous mice harboring this engineered allele (RMRP conditional).We found that insertion of this cassette suppressed RMRP expression, and we failed to obtain viable mice homozygous for the RMRP conditional allele.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Dana-Farber Cancer Institute, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, United States of America.

ABSTRACT
RMRP is a non-coding RNA that is ubiquitously expressed in both humans and mice. RMRP mutations that lead to decreased RMRP levels are found in the pleiotropic syndrome Cartilage Hair Hypoplasia. To assess the effects of deleting RMRP, we engineered a targeting vector that contains loxP sequences flanking RMRP and created hemizygous mice harboring this engineered allele (RMRP conditional). We found that insertion of this cassette suppressed RMRP expression, and we failed to obtain viable mice homozygous for the RMRP conditional allele. Furthermore, we were unable to obtain viable homozygous RMRP mice, indicating that RMRP is essential for early embryonic development.

Show MeSH
Related in: MedlinePlus