Generation of conditional knockout alleles for PRL-3

Hong Yan,Dong Kong,Xiaomei Ge,Xiang Gao,Xiao Han

doi:10.1016/s1674-8301(11)60058-4

Hong Yan, Dong Kong + Show 3 more

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https://doi.org/10.1016/s1674-8301(11)60058-4

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Abstract

Phosphatase of regenerating liver-3 (PRL-3) is a member of the protein tyrosine phosphatase (PTP) superfamily and is highly expressed in cancer metastases. For better understanding of the role of PRL-3 in tumor metastasis, we applied a rapid and efficient method for generating PRL-3 floxed mice and investigated its phenotypes. A BAC retrieval strategy was applied to construct the PRL-3 conditional gene-targeting vector. Exon 4 was selected for deletion to generate a nonfunctional prematurely terminated short peptide as it will cause a frame-shift mutation. Conditional knockout PRL-3 mice were generated by using the Cre-loxP system and were validated by Southern blot and RT-PCR analysis. Further analysis revealed the phenotype characteristics of PRL-3 knockout mice and wildtype mice. In this study, we successfully constructed the PRL-3 conditional knockout mice, which will be helpful to clarify the roles of PRL-3 and the mechanisms in tumor metastasis.

Highlights

The majority of cancer related deaths are induced by tumor metastasis and recent oncologic researches have focused on the mechanisms of metastasis
We have reported that Phosphatase of regenerating liver-3 (PRL-3) promoted motility and metastasis of mouse melanoma cells[7]
PRL-3 has been shown to play an important role in promoting cell migration and tumor metastasis

Summary

Introduction

The majority of cancer related deaths are induced by tumor metastasis and recent oncologic researches have focused on the mechanisms of metastasis. Protein tyrosine phosphatases (PTPs) play a fundamental role in regulating diverse cellular processes, including cell proliferation, adhesion and migration[1]. Phosphatase of regenerating liver-3 (PRL-3) is a member of the protein tyrosine phosphatase (PTP) superfamily. 荭 These authors contributed to this work. *Corresponding author: Xiao Han, Ph.D, Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, 140 Hanzhong Road, Nanjing, Jiangsu 210029, China. Sequences, and three mouse PRL proteins contain a Cterminal consensus sequence for prenylation[2]. PRL-3 encodes a small, -20 kDa tyrosine phosphatase that is located at the cytoplasmic membrane when prenylated at its C-terminus; the unprenylated PRL-3 is shifted into the nucleus[3].

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