title: Ð¡Ð¾Ð·Ð´Ð°Ð½Ð¸Ðµ ÑÑÐµÐ´ÑÑÐ² Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¸Ð½Ð¶ÐµÐ½ÐµÑÐ¸Ð¸ ÑÑÐºÐ°ÑÐ¸Ð¾Ñ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð¼Ð°Ð»Ð¾ÑÐ°Ð·Ð¼ÐµÑÐ½ÑÑ Cas9 Ð½ÑÐºÐ»ÐµÐ°Ð·
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 of eukaryotic genome engineering tools based on Cas9 nucleases of small-size
Creators: name: ÐÐ±ÑÐ°Ð¼Ð¾Ð²Ð°, ÐÐ°Ñ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: 2021
Subjects: ÐÐ»ÐµÑÐºÐ° (Ð±Ð¸Ð¾Ð».)
ÐÐ¸Ð¾ÑÐµÑ Ð½Ð¾Ð»Ð¾Ð³Ð¸Ñ
ÐÐ¼Ð¼ÑÐ½Ð¾Ð»Ð¾Ð³Ð¸Ñ
ÑÐµÐ´Ð°ÐºÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð³ÐµÐ½Ð¾Ð¼Ð°
genome editing

description: ÐÐ°Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð¿Ð¾ÑÐ²ÑÑÐµÐ½Ð° ÑÐ¾Ð·Ð´Ð°Ð½Ð¸Ñ ÑÑÐµÐ´ÑÑÐ² Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¸Ð½Ð¶ÐµÐ½ÐµÑÐ¸Ð¸ ÑÑÐºÐ°ÑÐ¸Ð¾Ñ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð½ÑÐºÐ»ÐµÐ°Ð· PpCas9 Ð¸Ð· Pasteurella pneumotropica Ð¸ DfCas9 Ð¸Ð· Defluviimonas sp.20V17. Ð ÑÐ¾Ð´Ðµ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¸ ÑÐ¾Ð·Ð´Ð°Ð½Ñ ÑÑÐºÐ°ÑÐ¸Ð¾ÑÐ¸ÑÐµÑÐºÐ¸Ðµ Ð²ÐµÐºÑÐ¾ÑÑ, Ð½ÐµÑÑÑÐ¸Ðµ CRISPR-PpCas9 Ð¸ CRISPR-DfCas9 ÑÐ¸ÑÑÐµÐ¼Ñ, Ð±ÑÐ»Ð° Ð¾ÑÑÑÐµÑÑÐ²Ð»ÐµÐ½Ð° Ð¿ÑÐ¾Ð²ÐµÑÐºÐ° Ð½ÑÐºÐ»ÐµÐ°Ð·Ð½Ð¾Ð¹ Ð°ÐºÑÐ¸Ð²Ð½Ð¾ÑÑÐ¸ Ð¸Ð·ÑÑÐ°ÐµÐ¼ÑÑ ÑÐ¸ÑÑÐµÐ¼ Ð² ÐºÐ»ÐµÑÐºÐ°Ñ ÑÐµÐ»Ð¾Ð²ÐµÐºÐ° HEK293T. CRISPR-DfCas9 ÑÐ¸ÑÑÐµÐ¼Ð° Ð½Ðµ Ð¿Ð¾ÐºÐ°Ð·Ð°Ð»Ð° ÑÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾Ð³Ð¾ ÑÐµÐ´Ð°ÐºÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð³ÐµÐ½Ð¾Ð¼Ð° ÐºÐ»ÐµÑÐ¾Ðº, Ð² ÑÐ¾ Ð²ÑÐµÐ¼Ñ ÐºÐ°Ðº CRISPR-PpCas9 Ð¾ÐºÐ°Ð·Ð°Ð»Ð°ÑÑ ÑÐ¿Ð¾ÑÐ¾Ð±Ð½Ð° Ð²Ð½Ð¾ÑÐ¸ÑÑ Ð´Ð²ÑÐ½Ð¸ÑÐµÐ²ÑÐµ ÑÐ°Ð·ÑÑÐ²Ñ Ð² ÐÐÐ ÐºÐ»ÐµÑÐ¾Ðº ÑÐµÐ»Ð¾Ð²ÐµÐºÐ°. Ð­ÑÑÐµÐºÑÐ¸Ð²Ð½Ð¾ÑÑÑ ÑÐ°Ð±Ð¾ÑÑ CRISPR-PpCas9 ÑÐ¸ÑÑÐµÐ¼Ñ Ð±ÑÐ»Ð° Ð¿Ð¾Ð²ÑÑÐµÐ½Ð° Ð¿ÑÑÐµÐ¼ Ð¿Ð¾Ð´Ð±Ð¾ÑÐ° Ð¾Ð¿ÑÐ¸Ð¼Ð°Ð»ÑÐ½Ð¾Ð¹ Ð´Ð»Ð¸Ð½Ñ ÑÐ¿ÐµÐ¹ÑÐµÑÐ½Ð¾Ð¹ ÑÐ°ÑÑÐ¸ Ð½Ð°Ð¿ÑÐ°Ð²Ð»ÑÑÑÐ¸Ñ Ð ÐÐ. ÐÐ»Ñ Ð´Ð¾ÑÑÐ°Ð²ÐºÐ¸ CRISPR-PpCas9 Ð² Ð¶Ð¸Ð²ÑÐµ Ð¾ÑÐ³Ð°Ð½Ð¸Ð·Ð¼Ñ Ð² Ð²Ð¸Ð´Ðµ AAV Ð¼Ñ ÑÐ¼Ð¾Ð³Ð»Ð¸ Ð¼Ð°ÐºÑÐ¸Ð¼Ð°Ð»ÑÐ½Ð¾ ÑÐ¼ÐµÐ½ÑÑÐ¸ÑÑ ÑÐ°Ð·Ð¼ÐµÑ Ð²ÐµÐºÑÐ¾ÑÐ½Ð¾Ð¹ ÐºÐ¾Ð½ÑÑÑÑÐºÑÐ¸Ð¸, Ð½ÐµÑÑÑÐµÐ¹ ÑÐ¸ÑÑÐµÐ¼Ñ, Ð¸ ÑÐ¿Ð°ÐºÐ¾Ð²Ð°ÑÑ ÐµÐµ Ð² Ð²Ð¸ÑÑÑÐ½ÑÐµ ÐºÐ°Ð¿ÑÐ¸Ð´Ñ. Ð¢Ð°ÐºÐ¸Ð¼ Ð¾Ð±ÑÐ°Ð·Ð¾Ð¼, Ð² ÑÐµÐ·ÑÐ»ÑÑÐ°ÑÐµ Ð´Ð°Ð½Ð½Ð¾Ð¹ ÑÐ°Ð±Ð¾ÑÑ Ð±ÑÐ»Ð¾ ÑÐ¾Ð·Ð´Ð°Ð½Ð¾ Ð½Ð¾Ð²Ð¾Ðµ ÑÑÐµÐ´ÑÑÐ²Ð¾ Ð³ÐµÐ½ÐµÑÐ¸ÑÐµÑÐºÐ¾Ð¹ Ð¸Ð½Ð¶ÐµÐ½ÐµÑÐ¸Ð¸ ÑÑÐºÐ°ÑÐ¸Ð¾Ñ Ð½Ð° Ð¾ÑÐ½Ð¾Ð²Ðµ Ð½ÑÐºÐ»ÐµÐ°Ð·Ñ PpCas9 Ð¸Ð· Pasteurella pneumotropica â Ð¼Ð°Ð»Ð¾ÑÐ°Ð·Ð¼ÐµÑÐ½Ð°Ñ ÑÐ¸ÑÑÐµÐ¼Ð°, Ð½Ð°Ð¿ÑÐ°Ð²Ð»ÐµÐ½Ð½Ð¾ Ð²Ð½Ð¾ÑÑÑÐ°Ñ ÑÐ°Ð·ÑÑÐ²Ñ Ð² Ð³ÐµÐ½Ð¾Ð¼Ð½ÑÑ ÐÐÐ ÐºÐ»ÐµÑÐ¾Ðº ÑÐµÐ»Ð¾Ð²ÐµÐºÐ°.
description: This work is devoted to the development of genome editing tools based on the nucleases PpCas9 from Pasteurella pneumotropica and DfCas9 from Defluviimonas sp. 20V17. In the course of the work, eukaryotic vectors carrying CRISPR-PpCas9 and CRISPR-DfCas9 systems were created, and the nuclease activity of PpCas9 and DfCas9 was tested in HEK293T human cells. The CRISPR-DfCas9 system did not demonstrate genome modification, while CRISPR-PpCas9 was able to introduce double-strand breaks in the genomic DNA. To improve the efficiency of the CRISPR-PpCas9 system we selected the optimal length of the spacer segment of the guide RNAs. To deliver CRISPR-PpCas9 to living organisms in the AAV form, we minimized the size of the vector carrying the system and package it in viral capsids. As a result of this work, a new tool for eukaryotic genome editing was created. It is based on the CRISPR-PpCas9 system from Pasteurella pneumotropica â a small-sized system that directionally introduces breaks in the genomic DNA of human cells.