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Related Concept Videos

DNA Topoisomerases02:02

DNA Topoisomerases

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Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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DNA replication involves the separation of the two strands of the double helix, with each strand serving as a template from which the new complementary strand is copied.  After replication, each double-stranded DNA includes one parental or “old” strand and one “new” strand. This is known as semiconservative replication. The resulting DNA molecules have the same sequence and are divided equally into the two daughter cells.
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DNA-only Transposons02:57

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
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DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation
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DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation

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DNA Transfection by Electroporation.

Priti Kumar, Arvindhan Nagarajan, Pradeep D Uchil

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    |July 3, 2019
    PubMed
    Summary
    This summary is machine-generated.

    Electroporation uses electrical fields to deliver DNA into cells, proving effective for cell lines resistant to other methods. Optimizing conditions is key for successful DNA delivery into new cell types.

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    Area of Science:

    • Molecular Biology
    • Biotechnology
    • Cell Biology

    Background:

    • Electroporation is a physical method for gene transfer into cells using electrical pulses.
    • It is effective for cell lines resistant to chemical or viral transfection methods like lipofection.
    • Optimization of electrical parameters is crucial for successful DNA delivery across different cell types.

    Purpose of the Study:

    • To describe a method for electroporation of DNA into mammalian cell lines.
    • To provide guidance on optimizing electroporation conditions for new cell lines.
    • To detail the use of the Gene Pulser Xcell Electroporation System for mammalian cell transfection.

    Main Methods:

    • Utilizing pulsed electrical fields to create transient pores in cell membranes.
    • Employing the Gene Pulser Xcell Electroporation System (Bio-Rad) for DNA delivery.
    • Empirical determination of optimal voltage, pulse duration, and buffer conditions for specific cell lines.

    Main Results:

    • Electroporation successfully introduced DNA into various animal cells, plant cells, and bacteria.
    • Demonstrated efficacy in cell lines refractory to traditional transfection techniques.
    • Established a protocol for mammalian cell line electroporation using a specific commercial system.

    Conclusions:

    • Electroporation is a versatile and effective method for DNA delivery, especially for challenging cell lines.
    • Empirical optimization is essential for maximizing transfection efficiency.
    • The Gene Pulser Xcell system provides a reliable platform for mammalian cell electroporation.