Difference between revisions of "Part:BBa K2295002"

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Hypoxia induced factors (HIFs) are transcription factors responding to decreased oxigen levels in the cellular environment.
 
Hypoxia induced factors (HIFs) are transcription factors responding to decreased oxigen levels in the cellular environment.
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Hypoxia induced factor 1 alpha (HIF1A) is a protein constantly expressed in mammalian cells. Under normoxic conditions HIF1A is hydroxylated and is marked by the E3 ubiquitin ligase which leads to the degradation by the proteasome. However, in hypoxic conditions HIF1A is stabilized and can heterodimerize with HIF1B. Also HIF1A transcription is often significantly upregulated under hypoxic conditions. Under hypoxic conditions HIF1 can then bind to hypoxia response elements (HREs) (<partinfo>BBa_K2295003</partinfo>) in the nucleus and lead to the expression of the gene of interest (Ziello et al., 2007).
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Hypoxia-inducible factor 1-alpha (HIF1A) is a protein constantly expressed in mammalian cells. Under normoxic conditions HIF1A is hydroxylated and is marked by the E3 ubiquitin ligase which leads to the degradation by the proteasome. However, in hypoxic conditions HIF1A is stabilized and can heterodimerize with HIF1B. Also HIF1A transcription is often significantly upregulated under hypoxic conditions. Under hypoxic conditions HIF1 can then bind to hypoxia response elements (HREs) (<partinfo>BBa_K2295003</partinfo>) in the nucleus and lead to the expression of the gene of interest (Ziello et al., 2007).
  
 
===Western Blot against HIF1A===
 
===Western Blot against HIF1A===

Revision as of 14:39, 1 November 2017


HIF1A (Hypoxia-inducible factor one alpha)


Mechanism

Figure 1: Signaling pathway of HIF1A.

Hypoxia induced factors (HIFs) are transcription factors responding to decreased oxigen levels in the cellular environment. Hypoxia-inducible factor 1-alpha (HIF1A) is a protein constantly expressed in mammalian cells. Under normoxic conditions HIF1A is hydroxylated and is marked by the E3 ubiquitin ligase which leads to the degradation by the proteasome. However, in hypoxic conditions HIF1A is stabilized and can heterodimerize with HIF1B. Also HIF1A transcription is often significantly upregulated under hypoxic conditions. Under hypoxic conditions HIF1 can then bind to hypoxia response elements (HREs) (BBa_K2295003) in the nucleus and lead to the expression of the gene of interest (Ziello et al., 2007).

Western Blot against HIF1A

Figure 2: Transient introduction of artificial HIF1A in HIF1A shRNA2 knockdown HEK293T cell line.

Induction of the overexpressed HIF1A-HA can be observed in the last lane. Endogenous and artificial HIF1A were detected with HIF1A antibody; reintroduced HIF1A was detected via an HA-tag; GAPDH served as a loading control. Wild type (WT) and knockdown (HIF1A shRNA2) cells were transfected with CMV:HIF1A-HA one day prior to induction with CoCl2 to simulate hypoxia. Cells were harvested 24 hours after induction. Immunoblot assay was performed using antibodies against HIF1A, HA-tag and GAPDH.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 79
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 10
    Illegal AgeI site found at 1086
  • 1000
    COMPATIBLE WITH RFC[1000]