By David M. Langenau
This quantity makes a speciality of defining the original attributes of utilizing the zebrafish melanoma version for locating vital pathways and power drug pursuits for the remedy of human cancers. utilizing the zebrafish version, the quantity explores oncogene and tumor suppressor discovery, chemical genetic techniques, genomics, epigenetics, melanoma imaging, and cellphone transplantation. Contributed chapters come from the main trendy laboratories operating during this box, which supplies a distinct viewpoint on zebrafish versions from a large spectrum of the study group. additionally, the publication bargains a close research of the most up-tp-date learn within the zone for particular zebrafish melanoma versions, together with T cellphone leukemia, rhabdomyosarcoma, liver and pancreatic melanoma, cancer, neuroblastoma, germ mobilephone tumors, and malignant peripheral sheath tumors. A bankruptcy can also be devoted to the improvement and usage of alternative piscine versions of melanoma. The compilation of chapters within the quantity culminates right into a finished and definitive textual content on zebrafish and melanoma, offering a miles wanted source at the strong attributes of the zebrafish version system.
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Additional resources for Cancer and Zebrafish: Mechanisms, Techniques, and Models
G) Increased transgene expression is achieved by application of up to 5 UAS repeats, while a much higher copy number reduces transcription efﬁciency. Systems similar to Gal4/UAS include the Q-system (H) and the TrpR/tUAS system (I) composed of the transactivator (pink oval) QF/TrpR and the upstream activation sequence (yellow rectangle) QUAS/tUAS, respectively. The QF can be inhibited by QS (blue half circle). (J) As these systems do not cross-react they can be combined in one model to study cell non-autonomous effects for example by expressing Gal4 and QF under different cell type speciﬁc promoters to elucidate the effect of disabled Notch-signaling (through expression of dominant-negative suppressor of hairless, grey rectangle) in B-cells, and assess the effect on oncogenic behavior of T-cells.
Proposed a method for RMCE in zebraﬁsh through phiC31 . Using a tol2 construct they expressed attP ﬂanked GFP under a tissue speciﬁc promoter and could exchange it with an attB ﬂanked mCherry. A drawback however is the relatively low efﬁciency of phiC31 integrase in genomic approaches. Another approach described by Mosimann et al. aims to establish single-insertion zebraﬁsh lines that harbor only one attP site . 34 M. Mayrhofer and M. Mione They developed three functional lines with single attP integration sites of which they characterized two with homogenous gene expression.
Through injection of two different gRNAs and Cas9 mRNA even huge chromosomal deletions are possible . To reduce mutation events the knock-out efﬁciency can also be titrated through varying RNA concentrations . As the mutations are inherited to the next generation the CRISPR/Cas9 system allows to generate stable knock-out lines with a fast and easy protocol . While off-target effects have been found to appear in only few cases, still deeper investigation is required . However, the system displays variable efﬁciencies for different genomic targets with some genes yielding effects in <1 % of injected embryos [117, 118, 122].