닫기
TaKaRa
로그인  |   회원가입  |   ID/PW 찾기   |  제품상담문의  
제품명/제품코드
키워드검색
검색
닫기
  • 고객지원 
  • 업무안내│제품문의
  • 전화번호│02-2081-2510
  • Email     │support@takara.co.kr
  • 업무시간안내
  • [ 평　　일 ] 09 : 00 ~ 18 : 00 │ [ 점심시간 ] 12 : 00 ~ 13 : 00
  • 토·일요일, 공휴일은 휴무입니다.
Home > 전제품보기 > Epigenetics > miRNA 발현 · 정량 > Tet Inducible miRNA 발현 시스템

Tet Inducible miRNA 발현 시스템

-

제조사 제품코드 제품명 용량 가격
(부가세별도)		 비고 사용자매뉴얼
데이터가 없습니다

  • Inducible microRNA expression using Tet-On Advanced
  • Track your inducible miRNA expression with fluorescent protein markers
  • Very high fold-induction and low basal expression
The Mir-X Inducible miRNA Red & Green Systems exploit key elements of our tightly regulated and highly responsive Tet-On Advanced Expression System to provide on-demand, high-level expression of your selected miRNA and a bright red or green fluorescent protein marker.
How It Works…
Your miRNA sequence is embedded in the 3’ UTR of a marker protein mRNA, which is expressed from an inducible miRNA expression vector (pmRi-mCherry or pmRi-ZsGreen1). When cells expressing the Tet-On Advanced transactivator are transfected with the pmRi-mCherry or pmRi-ZsGreen1 vector, high levels of your inducible microRNA and either the mCherry or ZsGreen1 proteins are expressed in response to doxycycline (Dox; Figures 1 & 2). Once Dox is added to the cell culture medium, miRNA induction is very rapid and effective. We used modified Metridia secreted luciferase genes containing miRNA target sequences (RNAiMonitor) to measure the effects of miR-1 and miR-9 miRNA induction in MCF7 Tet-On Advanced cells (Figure 3). Inducing miR-1 and miR-9 expression in these cells with Dox resulted in >90% knockdown of luciferase activity compared to controls.
Red and Green Complete Systems
The complete Mir-X Inducible miRNA Systems provide the pTet-On Advanced Vector for expressing the Dox-responsive transactivator; an inducible miRNA expression vector (either pmRi-mCherry or pmRi-ZsGreen1; also available separately); two linear selectable markers (hygromycin and puromycin) for cotransfecting with the pmRi vectors in order to establish stable cell lines; and tetracycline-free Tet System Approved FBS (serum) for ensuring the full activity of Tet-based expression systems.


Figure 1. MicroRNA induction in the Mir-X Inducible System. The Tet-controlled transactivator, Tet-On Advanced, is a fusion protein derived from a mutant version of the E. coli Tet repressor protein, rTetR, joined to three minimal HSV VP16 transcription activation domains. In the presence of doxycycline (Dox), Tet-On Advanced binds to the inducible promoter, PTight, and elicits high levels of transcription of a composite mRNA that encodes a fluorescent protein (mCherry or ZsGreen1), and your miRNA sequence in its 3’ UTR.


Figure 2. Cells expressing pmRi transcription units fluoresce brightly. MCF-7 Tet-On Advanced cells expressing an miR-1 miRNA construct from the pmRi-mCherry and pmRi-ZsGreen1 vectors (Panel A), exhibit bright red and green fluorescence, respectively, in response to Dox (Panels B & C).


Figure 3. The miR-1 and miR-9 microRNAs effectively reduce expression of target sequence-bearing luciferase genes. The RNAiMonitor System was used to generate luciferase mRNAs that contained miR-1 or miR-9 cognate target sequences in their 3’ UTRs (3 each). The target mRNAs were constitutively expressed in MCF-7 Tet-On Advanced cells that were cotransfected with pmRi-ZsGreen1 vectors containing miR-1 or miR-9 miRNA inserts. The parent pmRi-ZsGreen1 vector provided a negative control for each experiment. Inducing miR-1 or miR-9 expression with Dox resulted in >90% reduction in normalized luciferase activity.
Applications
  • Inducible miRNA expression controlled by doxycycline
Components
Mir-X Inducible miRNA Systems (Red and Green)
  • pTet-On Advanced Vector
  • pmRi-mCherry Vector (Red system only) or pmRi-ZsGreen1 Vector (Green system only)
    • pTRE-Tight-Luc Vector
    • Linear Hygromycin Marker
    • Linear Puromycin Marker
  • Tet System Approved FBS, US-Sourced
pmRi-mCherry and pmRi-ZsGreen1 Vectors
  • pmRi-mCherry Vector or pmRi-ZsGreen1 Vector
    • pTRE-Tight-Luc Vector
    • Linear Hygromycin Marker
    • Linear Puromycin Marker

Keyword : miRNA,microRNA,Mir-X,MirX