Mello Biotechnology
 
작성일 : 13-01-07
[Mello Biotechnology] Natural miRNA Precursor
세포 내에서 성숙형이 되는 natural miRNA 전구체
 
Natural miRNA Precursor
세포 내에서 성숙형이 되는 natural miRNA 전구체
 
 
Natural miRNA 전구체는 processing 되지 않은 miRNA transcript 입니다. 화학 합성품이 아니기 때문에, 세포 도입을 안심하고 할 수 있습니다. 세포 도입 후, miRNA processing 복합체와 신속하게 결합하고 processing되고 성숙 miRNA가 됩니다.
Natural miRNA 전구체는 single stranded RNA molecules이며, target gene silencing을 위한 RISC(RNA-induced silencing
complex)의 수확에 필요한 헤어핀 구조도 가지고 있습니다. 합성miRNA 대신, natural miRNA 전구체를 실험에 이용하는 이점으로 세포 내에 존재하는 천연의 miRNA 전구체와 같은 것으로 사용할 수 있는 것을 들 수 있습니다. Natural miRNA 전구체에서 생성된 성숙 miRNA는,다양한 유전자의 silencing  및 다른 유전자의 전사 활성화를 수행하기 위해 다양한 세포 내 process의 연구에 유용합니다.
 
                               Note : Natural microRNA sequence is in red.
 
 
Natural microRNA precursors are microRNA transcripts that have not yet been processed into mature microRNA. These
microRNA precursors can be delivered safely into the cell, where the precursors readily associate with microRNA processing complexes, and be processed into mature microRNAs. These precursors are single stranded RNA molecules that contain
the hairpin construct necessary to be assembled into RNA-induced silencing complexes (RISCs) for target gene silencing.
These molecules are the closest representation to the microRNA precursors found in cells. Mature microRNA can then be
used to study a multitude of cellular processes based on their ability to silence as well as indirectly upregulate various
genes. The advantage of using Natural microRNA precursors versus synthetic mimics is the ability to use the same
microRNA precursors that are present naturally in the cellular environment.
 
 
 
 
 Base sequence of miRNA(miR-302 & 367)
 
 
Features
  • Natural : Natural microRNA precursors, not synthetic siRNA nor shRNA mimics
  • Quality : Microarray certified composition quality
  • Quantity : High quantity available for in-vivo research (HPLC)
  • Validated : Corresponding siRNA/shRNA mimics are validated by publications
  • Custom microRNA precursors available!
 
Natural microRNA precursors from Mello Biotechnology are transcribed from a proprietary microRNA plasmid.  This
plasmid has been sequenced to ensure correct sequence, and it is the only natural representation of cellular microRNA
available on the market.  Traditionally, only synthetic mimics of microRNAs have been available, however it is unknown
whether these microRNA mimics may interfere with exact function in natural cell state.  The clear advantage of using
Natural microRNA precursor is the ability for the researcher to study the true role of microRNA in the cell without the
worry of other variables that come with using altered forms of microRNA that is not naturally found in cells.  They
are better able to represent the in vivo environment of an ESC, providing a system more closely related to nature than is
possible using synthetic microRNA mimics.
 
 
 
Why Natural ?
 
 
Natural microRNA Precursor facilitates research of authentic microRNA function.  It does so by delivering natural
microRNA function through a natural biogenesis mechanism.  Natural microRNA then functions via translational
suppression, whereas synthetic shRNA or siRNA simply mimic this function via direct messenger RNA degradation. 
Natural microRNA precursors provide researchers with the ability to eliminate other variables that can be introduced by
using mimics.  Since our Natural miRNA precursor is a naturally occurring single-stranded form that contains the same
hairpin constructs seen in natural cellular counterparts, and thus can perform the same functions.
 
Our microRNA Precursor delivers its function through a pathway that proceeds naturally.  Natural microRNA Precursor may contain both unmatched sense and anti-sense microRNA that are not complimentary to each other, whereas synthetic
siRNA and shRNA mimics have perfectly matching sense and anti-sense.  As a result, the anti-sense in synthetic mimics is
different from natural anti-sense microRNA.  This difference often results in different targeting which can lead to
unexpected off targeting effects.
 
 
 
Purified vs. Cell Extract
 
 
 
Natural microRNA precursors are provided in two forms: Cell Extract and Purified. Although both contain ample amount of precursors for experimental use, each version provide different benefits.  The raw extract form contains the hairpin
constructs, as well as other total RNA molecules.  The plasmid, which contains a GFP reporter, can be used as a reporter
to identify transfected cells.  In addition, total RNA protects microRNA precursors from degradation by RNAses, providing
a more stable shelf life.  The Purified form is pure microRNA precursor, without any vectors or debris.  This ensures that
only microRNA precursor is present, hence minimizing experimental variables.  However, removal of other RNA contents
in cell extract may decrease the shelf-life of mircoRNA precursors, and increase their susceptibility to RNAse degradation
 
 
 
Quality & Stability
 
 
 
Natural microRNA precursors generated from specific microRNA plasmids undergo quality analysis to ensure sequence
integrity.  These mircoRNA precursors contain the same sequence as naturally occurring cellular precursors and can
therefore be used in experiments to learn about the direct effects of microRNA on cellular processes.  The precursors
have been isolated from dicer-negative microRNA expressing competent cells that are capable of expressing ample
amount of specific microRNA precursors.  Since cells are dicer-negative, they only produce miRNA precursors but not
mature microRNAs.  Multiple purification processes are performed to ensure a high concentration of precursors in its pure
state.
 
Single-stranded RNase first is introduced to digest any impurity RNAs.  Since microRNA precursors hairpins contain double-stranded regions, only undesired single stranded RNAs are removed.  The microRNA precursor size and purification is
confirmed by electrophoresis.
 
Finally, the exact sequence and presence of the microRNA precursors are microarray certified to ensure that only pure
population of specific natural microRNA precursors are present.
 
 
Reference ;
 
 1. Lin et al., (2010) MicroRNA miR-302 inhibits the tumorigenecity of human pluripotent stem cells by coordinate
    suppression of CDK2 and CDK4/6 cell cycle pathways. Cancer Res., 70: 9473-9482.
 2. Lin et al., (2011) Regulation of somatic cell reprogramming through inducible miR-302 expression.  Nucleic Acids
    Res., 39: 1054-1065.
 3. Anokye-Danso et al., (2011) Highly efficient miRNA-mediated reprogramming of mouse and human somatic cells to
    pluripotency. Cell Stem Cell, 8: 376-388.
 4. Subramanyam D et al., (2011) Multiple targets of miR-302 and miR-373 promote reprogramming of humanfibroblasts
    to induced pluripotent stem cells. Nature Biotechnology., 29: 443-448.
 5. Miyoshi N et al., (2011) Reprogramming of mouse and human cells to pluripotency using mature microRNAs. Cell
    Stem Cell., 8: 633-638.
 
 
 
Ordering informations ;
 
Catalog Number
Product Name
Size
M-410-302-EX
miR-302bcad Cell Extract
1.7mg, 6.8mg & 86mg
M-410-302-PF
miR-302bcad Purified
10nmol, 40nmol & 500nmol
M-410-367-EX
miR-302bcad/367 Cell Extract
1.7mg, 6.8mg & 86mg
M-410-367-PF
miR-302bcad/367 Purified
10nmol, 40nmol & 500nmol
 
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