title: Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐºÐ° Ð¸ ÑÐµÑÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ ÐºÐ°ÑÑÐµÑ Ð³ÐµÐ½Ð½Ð¾Ð¹ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð´Ð»Ñ ÑÐ¾Ð·Ð´Ð°Ð½Ð¸Ñ ÑÐ¸ÑÑÐµÐ¼Ñ Ð¸Ð½Ð´ÑÑÐ¸Ð±ÐµÐ»ÑÐ½Ð¾Ð¹ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð³Ð¸Ð±ÐµÐ»Ð¸ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð³ÐµÐ½Ð° Ð´Ð¸ÑÑÐµÑÐ¸Ð¹Ð½Ð¾Ð³Ð¾ ÑÐ¾ÐºÑÐ¸Ð½Ð°
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¼Ð°Ð³Ð¸ÑÑÑÐ°
titleType: Subtitle
title: Development and testing of gene expression cassettes to create an inducible cell death system based on the diphtheria toxin gene
Creators: name: ÐÐ°Ð¹ÑÐµÐ²Ð°, Ð¯Ð½Ð°
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2020
Subjects: Ð´Ð¸ÑÑÐµÑÐ¸Ð¹Ð½ÑÐ¹ ÑÐ¾ÐºÑÐ¸Ð½
ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ñ
ÐºÐ»Ð¾Ð½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ
diphtheric toxin
expression
cloning

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° Ð¾ÑÐµÐ½ÐºÐµ ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»Ð° Ð¸Ð½Ð´ÑÑÐ¸Ð±ÐµÐ»ÑÐ½ÑÑ ÑÐ¸ÑÑÐµÐ¼ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð³Ð¸Ð±ÐµÐ»Ð¸ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð³ÐµÐ½Ð° Ð´Ð¸ÑÑÐµÑÐ¸Ð¹Ð½Ð¾Ð³Ð¾ ÑÐ¾ÐºÑÐ¸Ð½Ð°, ÑÑÐ±ÑÐµÐ´Ð¸Ð½Ð¸ÑÐ° A. ÐÐ°Ð´Ð°ÑÐ¸, ÐºÐ¾ÑÐ¾ÑÑÐµ ÑÐµÑÐ°Ð»Ð¸ÑÑ Ð² ÑÐ¾Ð´Ðµ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ: 1) Ð Ð°Ð·ÑÐ°Ð±Ð¾ÑÐ°ÑÑ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸, ÑÐºÑÐ¿ÑÐµÑÑÐ¸ÑÑÑÑÐ¸Ðµ DTA; 2) ÐÑÐ¾Ð²ÐµÑÑÐ¸ ÑÑÐ°Ð½ÑÑÐµÐºÑÐ¸Ð¸ Ð´Ð°Ð½Ð½ÑÐ¼Ð¸ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸ÑÐ¼Ð¸; 3) ÐÐ·ÑÑÐ¸ÑÑ Ð´Ð¸Ð½Ð°Ð¼Ð¸ÐºÑ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð¼Ð°ÑÐºÐµÑÐ° ÑÑÐ°Ð½ÑÑÐµÐºÑÐ¸Ð¸ (GFP) Ð² Ð¼Ð¾Ð´ÐµÐ»ÑÐ½Ð¾Ð¹ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð»Ð¸Ð½Ð¸Ð¸ (HEK293), ÑÐµÐ¼ ÑÐ°Ð¼ÑÐ¼ Ð¾ÑÐµÐ½Ð¸Ð² ÑÑÐ½ÐºÑÐ¸Ð¾Ð½Ð°Ð»ÑÐ½Ð¾ÑÑÑ Ð¿Ð¾Ð»ÑÑÐµÐ½Ð½ÑÑ ÑÐ¸ÑÑÐµÐ¼ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð³Ð¸Ð±ÐµÐ»Ð¸. Ð Ð°Ð±Ð¾ÑÐ° Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° Ð½Ð° Ð±Ð°Ð·Ðµ ÐÐÐ Â«ÐÐ¸Ð¾ÐºÐ°Ð´Â», Ð³Ð´Ðµ ÑÐ¾Ð±Ð¸ÑÐ°Ð»Ð°ÑÑ Ð·Ð½Ð°ÑÐ¸ÑÐµÐ»ÑÐ½Ð°Ñ ÑÐ°ÑÑÑ ÑÐ°ÐºÑÐ¸ÑÐµÑÐºÐ¾Ð³Ð¾ Ð¼Ð°ÑÐµÑÐ¸Ð°Ð»Ð°: ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÑ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ Ð¼Ð°ÑÐºÐµÑÐ° ÑÑÐ°Ð½ÑÑÐµÐºÑÐ¸Ð¸ (GFP) Ð² Ð¼Ð¾Ð´ÐµÐ»ÑÐ½Ð¾Ð¹ ÐºÐ»ÐµÑÐ¾ÑÐ½Ð¾Ð¹ Ð»Ð¸Ð½Ð¸Ð¸ (HEK293). ÐÑÐ»Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ñ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð´Ð¸Ð½Ð°Ð¼Ð¸ÐºÐ¸ ÑÐºÑÐ¿ÑÐµÑcÐ¸Ð¸ GFP, Ð¿Ð¾ÐºÐ°Ð·ÑÐ²Ð°ÑÑÐ¸Ðµ Ð½Ð°Ð³Ð»ÑÐ´Ð½Ð¾, ÐºÐ°ÐºÐ¸Ð¼ Ð¾Ð±ÑÐ°Ð·Ð¾Ð¼ ÑÐµ Ð¸Ð»Ð¸ Ð¸Ð½ÑÐµ ÑÐ»ÐµÐ¼ÐµÐ½ÑÑ Ð¿Ð»Ð°Ð·Ð¼Ð¸Ð´Ñ Ð²Ð»Ð¸ÑÑÑ Ð½Ð° ÑÐ¸ÑÐ¾ÑÐ¾ÐºÑÐ¸ÑÐµÑÐºÑÑ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ Ð´Ð¸ÑÑÐµÑÐ¸Ð¹Ð½Ð¾Ð³Ð¾ ÑÐ¾ÐºÑÐ¸Ð½Ð°. ÐÐ½Ð°Ð»Ð¸Ð· Ð¿ÑÐ¾Ð²Ð¾Ð´Ð¸Ð»ÑÑ Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ Ð¿ÑÐ¾ÑÐ¾ÑÐ½Ð¾Ð³Ð¾ ÑÐ¸ÑÐ¾ÑÐ»ÑÐ¾ÑÐ¸Ð¼ÐµÑÑÐ°. Ð ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ Ð±ÑÐ»Ð¸ ÑÐ°Ð·ÑÐ°Ð±Ð¾ÑÐ°Ð½Ñ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¸Ðµ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸, ÑÐºÑÐ¿ÑÐµÑÑÐ¸ÑÑÑÑÐ¸Ðµ DTA, Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð° ÑÑÐ°Ð½ÑÑÐµÐºÑÐ¸Ñ Ð¸ Ð¿ÑÐ¾Ð°Ð½Ð°Ð»Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ð° Ð´Ð¸Ð½Ð°Ð¼Ð¸ÐºÐ° ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ GFP. ÐÐ° Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð¸Ð¸ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð½ÑÑ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¹ Ð½Ð° ÐÐÐ Â«ÐÐ¸Ð¾ÐºÐ°Ð´Â», Ð¼Ð¾Ð¶Ð½Ð¾ ÑÐ´ÐµÐ»Ð°ÑÑ Ð²ÑÐ²Ð¾Ð´, ÑÑÐ¾ Ð´Ð¸ÑÑÐµÑÐ¸Ð¹Ð½ÑÐ¹ ÑÐ¾ÐºÑÐ¸Ð½ Ð¾Ð±Ð»Ð°Ð´Ð°ÐµÑ Ð²ÑÑÐ¾ÐºÐ¾Ð¹ ÑÐ¾ÐºÑÐ¸ÑÐ½Ð¾ÑÑÑÑ, Ð¸Ð¼ÐµÐ½Ð½Ð¾ Ð¿Ð¾ÑÑÐ¾Ð¼Ñ Ð´Ð»Ñ Ð¿ÑÐ¸Ð¼ÐµÐ½ÐµÐ½Ð¸Ñ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ¸ÑÑÐµÐ¼Ñ Ð¸Ð½Ð´ÑÐºÑÐ¸Ð¸ ÑÑÐµÐ±ÑÐµÑÑÑ Ð±Ð¾Ð»ÐµÐµ ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾Ðµ Ð¸Ð½Ð³Ð¸Ð±Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ ÑÐºÑÐ¿ÑÐµÑÑÐ¸Ð¸ DTA.
description: This work is devoted to evaluating the functionality of inducible cell death systems based on the diphtheria toxin gene, subunit A. The research set the following goals: 1) Develop genetic constructs expressing DTA; 2) Carry out transfection these constructs; 3) To study the dynamics of expression of the transfection marker (GFP) in a model cell line (HEK293), thereby evaluating the functionality of the resulting cell death systems. The work was fulfilled on the premises of Â«BiocadÂ», which included collection of factual materials, i.e. results of expression of the transfection marker (GFP) in a model cell line (HEK293).Studies have been conducted on the dynamics of GFP expression, which clearly show how certain plasmid elements affect the cytotoxic activity of diphtheria toxin. The analysis was carried out using a flow cytometer. As a result, genetic constructs expressing DTA were developed, transfection was performed, and the dynamics of GFP expression was analyzed. Based on the research conducted at Â«BiocadÂ», it can be concluded that diphtheria toxin is highly toxic, which is why the use of this induction system requires more effective inhibition of DTA expression.