By: Ashley Mignogna

Introduction
Transcription factor SOX-9 is a protein that in humans is encoded by the

SOX9 gene Belongs to the SOX gene family, SOX genes encode a family of transcription factors that bind to the minor groove in DNA Sox genes are classified into groups. Sox genes from different groups share little similarity outside the DNA-binding domain. In mouse and human the members of the groups are:

SoxA: SRY SoxB1: SOX1, SOX2, SOX3 SoxB2: SOX14, SOX21 SoxC: SOX4, SOX11, SOX12 SoxD: SOX5, SOX6, SOX13 SoxE: SOX8, SOX9, SOX10 SoxF: SOX7, SOX17, SOX18 SoxG: SOX15 SoxH: SOX30

Introduction cont..
located on chromosome 17
5,401 base pairs long 3 exons and 2 introns

126 SNPs
Recognizes the sequence CCTTGAG

Functions
Transcription factor essential

for both sex and skeletal development. Other functions include regulatory elements, cancer, skin pigmentation, and organ maintenance

SOX-9 Reporter
Reporter genes are often used as an indication of whether a certain

gene has been taken up by or expressed in the cell or organism population Reporters are genes that researchers attach to a regulatory sequence of another gene of interest and are easy to locate. The reporter found in the sox-9 gene increases the activity of the promoter; this reporter is known as SOX9cre1. It is located 1.1Mb away from the Sox-9 gene as shown in the picture below.

Overexpression of SOX-9
Sox-9 has varying roles in many different tissues.

Overexpression of this gene causes different results in the tissues in which it resides. This overexpression can affect things anywhere from chondrocytes to heart valves. Overexpression of Sox9 in chondrocytes, cells found within cartilage, using Col2a1-Cre mice suppressed chondrocyte hypertrophy, and delayed terminal differentiation and subsequent ossification.
Hypertrophy- increased in cell volume

In another study it was found that overexpression of the SOX-9

gene can decrease the calcification process in the heart valves.

Calcification is the process in which calcium salts build up in soft

tissue, causing it to harden

SOX-9 Conditional Knockout

Conditional gene knockout is a technology in which a specific target

gene is eliminated from a single organ in the body In one study, a conditional knockout targeting vector of Sox9 was generated by flanking exon 1 with loxP sites. Homozygous mice for the Sox9 allele are viable, fertile and indistinguishable from wild type mice, indicating that the Sox9 allele is a fully functional Sox9 allele. Furthermore, they demonstrated that Cre-mediated recombination using a Col2a1-Cre line resulted in specific removal of Sox9 activity in cartilage tissues.

Conditional Knockout Cont..

In another study, to determine whether Sox9 is required for

testis development after the initial steps of sex determination, Sox9 mice were crossed with an AMH-Cre transgenic line thereby completely deleting Sox9 in Sertoli cell.
Sertoli cells: a 'nurse' cell of the testes that is part of a seminiferous

tubule. Its main function is to nourish the developing sperm cells through the stages of spermatogenesis.

Conditional Sox9 null mutants show normal embryonic

testis development and are initially fertile, become sterile from dysfunctional spermatogenesis at about 5 months. The mutant testis rapidly and progressively lost weight

Reduced Sox-9 Expression

Sox9 gene is also expressed in developing and mature heart valves.
It is required for expression of cartilage-associated proteins. Mice with

reduced sox9 function develop cartilage defects, and gain skeletal bone prematurely. It was found that from 3 months of age, Sox9(fl/+);Col2a1-cre mice develop calcification lesions in heart valve leaflets associated with increased expression of bone-related genes and activation of inflammation and matrix remodeling processes. Abnormal calcification is also observed following direct knockdown of Sox9 in heart valves in vitro.
Gene knockdown refers to techniques by which the expression of one or

more of an organism's genes is reduced, either through genetic modification or treatment with a reagent

Campomelic Displaysia
Genetic disorder refering to the bowing of the femur and

tibia. A mutation in the gene coding for the transcription factor Sox9 Symptoms:

irregularities of the ribs and vertebral column bowing of the long bones. short legs dislocated hips and underdeveloped shoulder blades 11 pairs of ribs instead of 12 bone abnormalities in the neck and feet that are abnormally rotated (clubfeet).

Campomelic Displaysia Cont..

Sox9 controls transcription of Type II collagen and the

proteoglycan aggrecan.
Type-II collagen is the basis for articular cartilage and

hyaline cartilage. Proteoglycan Aggrecan is an integral part of the extracellular matrix in cartilaginous tissue and it withstands compression in cartilage.

Sex reversal
Another part of the campomelic displaysia disorder

involving SOX9 is related to sex reversal. SOX9 is not able to fulfill its role in sex determination due to mutations around or in the gene. Genetically male individuals appear phenotypically female 75 percent of affected individuals with a typical male chromosome pattern have ambiguous genitalia or normal female genitalia
Internal reproductive organs may not correspond with the

external genitalia; they can be male, female, or a combination of the two.

Summary
It acts as a transcription factor during chondrocyte

differentiation Deficiencies lead to the skeletal malformation syndrome campomelic dysplasia, frequently with sex reversal. Plays an important role in the normal skeletal development. Regulates the expression of other genes