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Method for establishing drosophila melanogaster model for inducing colon cancer high-frequency metastasis

A technology of high-frequency transfer and method establishment, applied in the field of biogenetics, can solve the problems of inability to effectively simulate the dynamic characteristics of tumor metastasis in patients, unsuitable for drug screening and mechanism, short larval stage, etc., to achieve short reproductive cycle, in vivo tissue The effect of clear organ differentiation and perfect adult development

Active Publication Date: 2022-04-01
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are few tumor migration models in Drosophila, most of which are larval stage imago disc models, which are screened in the larval stage, which is limited by the short larval stage (about 5 days in larval stage), acute tumor metastasis, and difficult administration. Cannot effectively simulate the dynamic characteristics of patient tumor metastasis and the relationship between research and body aging
Most of the existing Drosophila adult tumor metastasis models are colon cancer, but the transfer efficiency is very low, which is not suitable for drug screening and mechanism research

Method used

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  • Method for establishing drosophila melanogaster model for inducing colon cancer high-frequency metastasis
  • Method for establishing drosophila melanogaster model for inducing colon cancer high-frequency metastasis
  • Method for establishing drosophila melanogaster model for inducing colon cancer high-frequency metastasis

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Embodiment 1

[0031] A method for establishing a Drosophila melanogaster model that induces high-frequency metastasis of colon cancer, such as figure 1 shown, including the following steps:

[0032] S1: set lgl - / - UASRas V12 , FRT40A / Cyo; TM3 / TM6B Drosophila strains and Sp / Cyo; UASRas V12 , Sna OE / TM3 / MKRS Drosophila strains were crossed, and the genotype was selected as lgl in the offspring - / - UASRas V12 , FRT40A / Cyo; UASRas V12 , Sna OE / TM6B, to establish a Drosophila strain that can be subcultured;

[0033] S2: The genotype selected in the Drosophila library is: hsflp; FRT40A, tubGal80 / Cyo; tubGal4, UASGFP / TM6B virgin flies and the genotype selected in S1 is lgl - / - UASRRasV12 , FRT40A / Cyo; UASRas V12 , UASSna OE / TM6B male fruit flies are crossed, and the genotype hsflp; lgl is selected in the offspring - / - UASRas V12 , FRT40A / FRT40A, tubGal80; UASRas V12 , UASSna OE / tubGal4, the female adult of UASGFP, its appearance phenotype is flat wings, and the number of shoulde...

Embodiment 2

[0050] A method for establishing a Drosophila melanogaster model that induces high-frequency metastasis of colon cancer. In order to correctly prepare the medium for raising Drosophila; all Drosophila strains involved in the experiment were raised in: sugar 100g, corn flour 320g, 40g of soybean powder, 260g of brown sugar syrup, 40g of agar, 72g of yeast powder, and 5L of water. Continuously weigh 100g of sugar, 320g of corn flour, and 40g of soybean flour; add 5L of water to the pot and heat to 40-50°C; add about 600ml of hot water to the beaker until the beaker is almost full, stir and mix well; Add the weighed agar and yeast powder, stirring constantly with a spoon; add the mixture in the beaker, stirring constantly. Then use a beaker to weigh 260g of brown sugar syrup and add it to the pot. The syrup remaining on the inner wall of the beaker can be added to the beaker with cold water and placed in a microwave oven for about 1 minute to dissolve and then added to the pot; k...

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Abstract

The invention discloses a method for establishing a drosophila melanogaster model for inducing colon cancer high-frequency metastasis, and relates to the technical field of biogenetics. The invention aims to establish tumor migration which is obvious in migration, high in migration efficiency and stable in adult body. The method specifically comprises the following steps: adding lgl <- / -> UASRasV12 and FRT40A / Cyo into a reaction kettle; a TM3 / TM6B fruit fly strain and Sp / Cyo; the method comprises the following steps: hybridizing fruit fly strains of lgl <-> / -UASRasV12 and SnaOE / TM < 3 > / MKRS, and selecting FRT40A / Cyo with genotypes of lgl <-> / -UASRasV12 and FRT40A / Cyo from later generations; according to the method, a fruit fly strain capable of being subjected to passage preservation is built by using a single-cell culture medium, UASRasV12 and SnaOE / TM6B. According to the invention, fruit fly colon cancer is taken as a model, and an efficient tumor metastasis model is established through genetic manipulation of expression of tumor related factors such as Ras and lgl, Sna and the like.

Description

technical field [0001] The invention relates to the technical field of biogenetics, in particular to a method for establishing a Drosophila melanogaster model that induces high-frequency metastasis of colon cancer. Background technique [0002] MARCM (Mosaic analysis with a repressible maker) is a technical tool for studying the genetic development of Drosophila combined with FLP / FRT and UAS / Gal4 / Gal80 systems. Using MARCM technology can enable Among them, the UAS / Gal4 system increases the expression of the target gene in the chimera, and realizes the expression of the gene we are interested in in a specific tissue (it can be mutated), and uses the FLP / FRT system to insert Gal80 on one arm of the normal chromosome to make the normal The cells cannot express fluorescence, and the mutant chromosomes delete Gal80 through homologous recombination, and Gal4 and UAS combine to express the target gene to obtain the desired chimeric cells. GFP-positive cells are observed under a flu...

Claims

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Application Information

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IPC IPC(8): A01K67/033
Inventor 陈纾昕孟天恺邓寒松
Owner TONGJI UNIV
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